Cephem compounds

ABSTRACT

Bacteriostatic cephem compounds and processes for preparing same having the formula: ##STR1## wherein R 1  is amino or protected amino, 
     R 2  is lower alkyl, amino-(lower)-alkyl, protected amino-(lower)-alkyl, hydroxy-(lower)-alkyl, protected hydroxy-(lower)-alkyl, lower alkylthio-(lower)-alkyl, carboxy-(lower)-alkyl, esterified carboxy-(lower)-alkyl, (C 3  to C 8 ) cycloalkyl, lower alkenyl or lower alkynyl, 
     R 3  is a heterocyclic group substituted with amino(lower)alkyl, protected amino(lower)-alkyl, hydroxy(lower)alkyl or both amino and lower alkyl, 
     R 4  carboxy or protected carboxy, 
     provided that R 3  is a heterocyclic group substituted with hydroxy(lower)alkyl or both amino and lower alkyl, when R 2  is lower alkyl, 
     and its pharmaceutically acceptable salt.

This is a continuation-in-part of copending application, Ser. No.941,660, filed Sept. 12, 1978, now U.S. Pat. No. 4,220,761.

This invention relates to new cephem compound. More particularly, itrelates to new 3,7-disubstituted-3-cephem-4-carboxylic acid and itspharmaceutically acceptable salt, which have antimicrobial activities,and processes for preparation thereof, to pharmaceutical compositioncomprising the same and method of using the same prophylactically and atherapeutically for treatment of infectious diseases in human being andanimals.

Accordingly, the objects of this invention are to provide:

new 3,7-disubstituted-3-cephem-4-carboxylic acid and itspharmaceutically acceptable salt, which exhibit excellent antimicrobialactivities against a wide variety of pathogenic microorganisms includingGram negative and Gram positive bacteria,

processes for preparation of the same,

pharmaceutical composition comprising one of the same as an activeingredient, and

a method of using the same prophylactically and therapeutically fortreatment of infectious diseases caused by pathogenic microorganisms inhuman being and animals.

The cephem compound provided by this invention can be represented by theformula (I): ##STR2## wherein

R¹ is amino or protected amino,

R² is lower alkyl, amino-(lower)-alkyl, protected amino-(lower)-alkyl,hydroxy-(lower)-alkyl, protected hydroxy-(lower)-alkyl, loweralkylthio-(lower)-alkyl, carboxy-(lower)-alkyl, esterifiedcarboxy-(lower)-alkyl, (C₃ to C₈) cycloalkyl, lower alkenyl or loweralkynyl,

R³ is a heterocyclic group substituted with amino(lower)alkyl, protectedamino(lower)-alkyl, hydroxy(lower)alkyl or both amino and lower alkyl,

R⁴ carboxy or protected carboxy,

provided that R³ is a heterocyclic group substituted withhydroxy(lower)alkyl or both amino and lower alkyl, when R² is loweralkyl, and its pharmaceutically acceptable salt.

The compound (I) of this invention can be prepared by processes as shownin the following scheme. ##STR3## wherein

R¹, R², R³ and R⁴ are each as defined above,

R_(a) ¹ is protected amino,

R_(a) ² is protected hydroxy(lower)alkyl,

R_(b) ² is hydroxy(lower)alkyl,

R_(c) ² is protected amino(lower)alkyl,

R_(d) ² is amino(lower)alkyl,

R_(e) ² is esterified carboxy(lower)alkyl,

R_(f) ² is carboxy(lower)alkyl,

R_(a) ³ is heterocyclic group substituted with protectedamino(lower)alkyl,

R_(b) ³ is heterocyclic group substituted with amino(lower)alkyl,

R_(a) ⁴ is protected carboxy, and

R⁵ is acyl.

Among the starting compound in the present invention, some of thestarting compound (III) used in Process A are novel and can berepresented by the following formula: ##STR4## wherein

R¹ is amino or protected amino, and

R_(g) ² is amino(lower)alkyl, lower alkoxycarbonylamino(lower)alkyl,hydroxy(lower)alkyl, protected hydroxy(lower)alkyl, loweralkylthio(lower)alkyl, carboxy(lower)alkyl, esterifiedcarboxy(lower)alkyl, (C₃ to C₈) cycloalkyl or lower alkynyl, and itsester and a salt thereof.

The starting compound (IIIa) can be prepared by the methods illustratedbelow. ##STR5## where

R¹, R_(a) ¹, R_(a) ², R_(b) ², R_(d) ² and R_(g) ² are each as definedabove,

R_(h) ² is lower alkoxycarbonylamino(lower)alkyl

R_(i) ² is azido(lower)alkyl,

X is halogen, and

Z is esterified carboxy.

The terms and definitions described in this specification areillustrated as follows.

(a) Partial structure of the formula: ##STR6## is intended to mean bothof the geometric formulae: ##STR7##

From the view point of structure-activity relationship, it is to benoted that a syn isomer of the compound (I) tends to be of much higherantimicrobial activity than the corresponding anti isomer, andaccordingly the syn isomer of the compound (I) is more preferableantimicrobial agent than the corresponding anti isomer in theprophylactic and therapeutic value.

(b) The thiazolyl group of the formula: ##STR8## (wherein R¹ is asdefined above) is well known to lie in tautomeric relation with athiazolinyl group, and the tautomerism between the said thiazolyl andthiazolinyl groups can be illustrated by the following equilibrium:##STR9## (wherein R¹ is as defined above, and R^(1') is imino orprotected imino)

Accordingly, it is to be understood that both of the said groups aresubstantially the same, and the tautomers consisting of such groups areregarded as the same compounds. Therefore, both of the tautomeric formsof the compounds having such groups in their molecule are included inthe scope of this invention and designated inclusively with oneexpression "thiazolyl" and represented by the formula: ##STR10##(wherein R¹ is as defined above) only for the convenient sake throughoutthis specification.

In the above and subsequent descriptions of this specification, suitableexamples and illustration of the various definitions which thisinvention intends to include within the scope thereof are explained indetail as follows.

The term "lower" is used to intend a group having 1 to 6 carbon atom(s),unless otherwise provided.

The term "protective group" in the terms "protected amino" and"protected amino(lower)alkyl" may include a conventional N-protectivegroup such as acyl, substituted or unsubstituted ar(lower)-alkyl (e.g.benzyl, benzhydryl, trityl, 4-methoxybenzyl, 3,4-dimethoxybenzyl, etc.),halo(lower)alkyl (e.g. trichloromethyl, trichloroethyl, trifluoromethyl,etc.), tetrahydropyranyl, substituted phenylthio, substitutedalkylidene, substituted aralkylidene, substituted cycloalkylidene, orthe like.

Suitable acyl for the N-protective group may be aliphatic acyl group andacyl group containing an aromatic or heterocyclic ring. And, suitableexamples of the said acyl may be lower alkanoyl (e.g., formyl, acetyl,propionyl, butyryl, isobutyryl, valeryl, isovaleryl, oxalyl, succinyl,pivaloyl, etc.), preferably one having 1 to 4 carbon atom(s), morepreferably one having 1 to 2 carbon atom(s); lower alkoxycarbonyl having2 to 7 carbon atoms (e.g., methoxycarbonyl, ethoxycarbonyl,propoxycarbonyl, 1-cyclopropylethoxycarbonyl, isopropoxycarbonyl,butoxycarbonyl, t-butoxycarbonyl, pentyloxycarbonyl,t-pentyloxycarbonyl, hexyloxycarbonyl, etc.), preferably one having 2 to6 carbon atoms; lower alkanesulfonyl (e.g., mesyl, ethanesulfonyl,propanesulfonyl, isopropanesulfonyl, butanesulfonyl, etc.);arenesulfonyl (e.g., benzenesulfonyl, tosyl, etc.); aroyl (e.g.,benzoyl, toluoyl, naphthoyl, phthaloyl, indancarbonyl, etc.);ar(lower)alkanoyl (e.g., phenylacetyl, phenylpropionyl, etc.);ar(lower)alkoxycarbonyl (e.g., benzyloxycarbonyl, phenethyloxycarbonyl,etc.); and the like.

The acyl as stated above may have 1 to 3 suitable substituent(s) such ashalogen (e.g., chlorine, bromine, iodine or fluorine), hydroxy, cyano,nitro, lower alkoxy (e.g., methoxy, ethoxy, propoxy, isopropoxy, etc.),lower alkyl (e.g., methyl, ethyl, propyl, isopropyl, butyl, etc.), loweralkenyl (e.g., vinyl, allyl, etc.), aryl (e.g., phenyl, tolyl, etc.), orthe like, and preferable example is mono(or di ortri)halo(lower)alkanoyl (e.g., chloroacetyl, dichloroacetyl,trifluoroacetyl, etc.).

And further, the reaction product of a silan, boron, aluminium orphosphorus compound with the amino group may also be included in theN-protective group. Suitable examples of such compounds may betrimethylsilyl chloride, trimethoxysilyl chloride, boron trichloride,butoxyboron dichloride, aluminum trichloride, diethoxy aluminumchloride, phosphorus dibromide, phenylphosphorus dibromide, or the like.

The term "lower alkyl" and "lower alkyl moiety" in the terms"amino(lower)alkyl", "protected amino(lower)alkyl","hydroxy(lower)alkyl", "protected hydroxy(lower)alkyl", "loweralkylthio(lower)alkyl", "carboxy(lower)alkyl", "esterifiedcarboxy(lower)alkyl" and "lower alkoxycarbonylamino(lower)alkyl" mayinclude a residue of straight and branched alkane having 1 to 6 carbonatom(s) such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl,t-butyl, pentyl, neopentyl, hexyl and the like, and preferably the onehaving 1 to 4 carbon atom(s).

The term "protective group" in the term "protected hydroxy(lower)alkyl"may include a conventional O-protective group such as acyl asaforementioned, or the like.

The term "(C₃ to C₈)cycloalkyl" may include cyclopropyl, cyclopentyl,cyclohexyl, cycloheptyl and the like, and preferably (C₅ toC₆)cycloalkyl.

The term "lower alkenyl" may include a residue of a straight or branchedalkene having 2 to 6 carbon atoms, such as vinyl, allyl, 1-propenyl,isopropenyl, butenyl, isobutenyl, pentenyl, hexenyl and the like, andpreferably the ones having up to 5 carbon atoms.

The term "lower alkynyl" may include a residue of a straight or branchedalkyne having 2 to 6 carbon atoms, such as ethynyl, propargyl,1-propynyl, 3-butynyl, 2-butynyl, 4-pentynyl, 3-pentynyl, 2-pentynyl,1-pentynyl, 5-hexynyl and the like, and preferably the ones having up to5 carbon atoms.

The term "heterocyclic group" in the term "a heterocyclic groupsubstituted with amino(lower)alkyl, protected amino(lower)alkyl,hydroxy(lower)alkyl or both amino and lower alkyl" may includeunsaturated 5 to 6-membered heterocyclic group containing at least onehetero atom selected from oxygen, sulfur and nitrogen atoms.

And, preferable heterocyclic group may be the one such as

unsaturated 5 to 6-membered heterocyclic group containing 1 to 2 oxygenatom(s), for example, furyl;

unsaturated 5 to 6-membered heterocyclic group containing 1 to 4nitrogen atom(s), for example, pyrrolyl, imidazolyl, pyrazolyl, pyridyl,picolyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazolyl (e.g.,4H-1,2,4-triazolyl, 1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, etc.),tetrazolyl (e.g., 1H-tetrazolyl, 2H-tetrazolyl, etc.), etc.;

unsaturated 5 to 6-membered heterocyclic group containing 1 to 2 oxygenatom(s) and 1 to 3 nitrogen atom(s), for example, oxazolyl, isoxazolyl,oxadiazolyl (e.g., 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl,1,2,5-oxadiazolyl, etc.), etc.;

unsaturated 5 to 6-membered heterocyclic group containing 1 to 2 sulfuratom(s) and 1 to 3 nitrogen atom(s), for example, thiazolyl,isothiazolyl, thiadiazolyl (e.g., 1,2,4-thiadiazolyl,1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, etc.), etc.; and the like.

The term "protected carboxy" may include esterified carboxy, amidatedcarboxy or the like.

Suitable examples of "the ester" and "ester moiety" in the "esterifiedcarboxy" may be lower alkyl ester (e.g., methyl ester, ethyl ester,propyl ester, isopropyl ester, butyl ester, isobutyl ester, t-butylester, pentyl ester, t-pentyl ester, hexyl ester, 1-cyclopropylethylester, etc.);

lower alkenyl ester (e.g., vinyl ester, allyl ester, etc.);

lower alkynyl ester (e.g., ethynyl ester, propynyl ester, etc.);

lower alkoxy(lower)alkyl ester (e.g., methoxymethyl ester, ethoxymethylester, isopropoxymethyl ester, 1-methoxyethyl ester, 1-ethoxyethylester, etc.);

lower alkylthio(lower)alkyl ester (e.g., methylthiomethyl ester,ethylthiomethyl ester, ethylthioethyl ester, isopropylthiomethyl ester,etc.);

halo(lower)alkyl ester (e.g., 2-iodoethyl ester, 2,2,2-trichloroethylester, etc.);

lower alkanoyloxy(lower)alkyl ester (e.g., acetoxymethyl ester,propionyloxymethyl ester, butyryloxymethyl ester, valeryloxymethylester, pivaloyloxymethyl ester, hexanoyloxymethyl ester, 2-acetoxyethylester, 2-propionyloxyethyl ester, etc.);

lower alkanesulfonyl(lower)alkyl ester (e.g., mesylmethyl ester,2-mesylethyl ester, etc.);

ar(lower)alkyl, for example, phenyl(lower)alkyl ester which may have oneor more suitable substituent(s) (e.g., benzyl ester, 4-methoxybenzylester, 4-nitrobenzyl ester, phenethyl ester, trityl ester, benzhydrylester, bis(methoxyphenyl)methyl ester, 3,4-dimethoxybenzyl ester,4-hydroxy-3,5-di-t-butylbenzyl ester, etc.);

aryl ester which may have one or more suitable substituent(s) (e.g.,phenyl ester, tolyl ester, t-butylphenyl ester, xylyl ester, mesitylester, cumenyl ester, salicyl ester, etc.); an ester with a silylcompound such as tri(lower)-alkylsilyl compound,di(lower)alkylalkoxysilyl compound or tri(lower)alkoxysilyl compound,for example, tri(lower)alkylsilyl ester (e.g., trimethyl silyl ester,triethylsilyl ester, etc.), di(lower)alkylalkoxy silyl ester (e.g.,dimethylmethoxysilyl ester, dimethylethoxysilyl ester,diethylmethoxysilyl ester, etc.) or tri(lower)-alkoxysilyl ester (e.g.,trimethoxysilyl ester, triethoxysilyl ester, etc.), or the like.

More particularly, the preferable example of ester may benitrophenyl(lower)alkyl ester (e.g., 4-nitrobenzyl ester,4-nitrophenethyl ester, etc.), lower alkyl ester (e.g., methyl ester,ethyl ester, propyl ester, isopropyl ester, butyl ester, isobutyl ester,tert-butyl ester, pentyl ester, neopentyl ester, hexyl ester, etc.).

The term "esterified carboxy moiety" in the term "esterifiedcarboxy(lower)alkyl" may be referred to the aforementioned examples ofesterified carboxy.

More particularly, the preferable examples of R¹ to R⁴ are illustratedas follows.

The preferable examples of R¹ may be amino or acylamino [morepreferably, lower alkanoylamino (e.g., formamido, acetamido, etc.) ortrihalo(lower)alkanoylamino (e.g., trifluoroacetamido, etc.)].

The preferable examples of R² may be lower alkyl (e.g., methyl, ethyl,propyl, isopropyl, butyl, isobutyl, pentyl, hexyl, etc.),amino(lower)alkyl (e.g., aminomethyl, aminoethyl, aminopropyl, etc.),acylamino(lower)alkyl [more preferably, loweralkoxycarbonylamino(lower)alkyl (e.g., methoxycarbonylaminomethyl,ethoxycarbonylaminoethyl, propoxycarbonylaminopropyl,t-butoxycarbonylaminoethyl, t-butoxycarbonylaminopropyl, etc.)],hydroxy(lower)alkyl (e.g., hydroxymethyl, hydroxyethyl, hydroxypropyl,etc.), acyloxy(lower)alkyl [more preferabl, loweralkanoyloxy(lower)alkyl (e.g., formyloxymethyl, formyloxyethyl,acetoxyethyl, formyloxypropyl, etc.)], lower alkylthio(lower)alkyl(e.g., methylthiomethyl, methylthioethyl, ethylthioethyl, etc.),carboxy(lower)alkyl (e.g., carboxymethyl, carboxypropyl, etc.),esterified carboxy(lower)alkyl [more preferably, loweralkoxycarbonyl(lower)alkyl (e.g., methoxycarbonylmethyl,ethoxycarbonylmethyl, t-butoxycarbonylmethyl, propoxycarbonylethyl,etc.)], (C₃ to C₈)cycloalkyl [more preferably, (C₅ to C₆)cycloalkyl(e.g., cyclopentyl, cyclohexyl, etc.)], lower alkenyl (e.g., vinyl,allyl, etc.) or lower alkynyl (e.g., ethynyl, propargyl, etc.).

The preferable examples of R³ may be

unsaturated 5 to 6-membered heterocyclic group containing 1 to 4nitrogen atom(s) substituted with amino(lower)alkyl, loweralkoxycarbonylamino(lower)alkyl, hydroxy(lower)alkyl or both amino andlower alkyl [more preferably, amino(lower)alkyltetrazolyl (e.g.,1-aminomethyl-1H-tetrazol-5-yl, 1-(2-aminoethyl)-1H-tetrazol-5-yl,1-(3-aminopropyl)-1H-tetrazol-5-yl, etc.), loweralkoxycarbonylamino(lower)alkyltetrazolyl (e.g.,1-methoxycarbonylaminomethyl-1H-tetrazol-5-yl,1-(2-t-butoxycarbonylaminoethyl)-1H-tetrazol-5-yl,1-(3-t-butoxycarbonylaminopropyl)-1H-tetrazol-5-yl, etc.),hydroxy(lower)alkyltetrazolyl (e.g., 1-hydroxymethyl-1H-tetrazol-5-yl,1-(2-hydroxyethyl)-1H-tetrazol-5-yl,1-(3-hydroxypropyl)-1H-tetrazol-5-yl, etc.) or triazolyl substitutedwith both amino and lower alkyl (e.g.,4-amino-5-methyl-4H-1,2,4-triazol-3-yl,4-amino-5-ethyl-4H-1,2,4-triazol-3-yl, etc.)] or

unsaturated 5 to 6-membered heterocyclic group containing 1 to 2 sulfuratom(s) and 1 to 3 nitrogen atom(s) substituted with amino(lower)alkylor lower alkoxycarbonylamino(lower)alkyl [more preferably,amino(lower)alkylthiadiazolyl (e.g.,5-aminomethyl-1,3,4-thiadiazol-2-yl,5-(2-aminoethyl)-1,3,4-thiadiazol-2-yl, etc.) or loweralkoxycarbonylamino (lower)alkylthiadiazolyl (e.g.5-methoxycarbonylaminomethyl-1,3,4-thiadiazol-2-yl,5-t-butoxycarbonylaminomethyl-1,3,4-thiadiazol-2-yl,5-(2-t-butoxycarbonylaminoethyl)-1,3,4-thiadiazol-2-yl, etc.)].

The preferable example of R⁴ may be carboxy.

Suitable "pharmaceutically acceptable salt" of the compound (I) includesa conventional non-toxic salt, and may be a salt with an inorganic baseor acid, for example, a metal salt such as an alkali metal salt (e.g.sodium salt, potassium salt, etc.) and an alkaline earth metal salt(e.g. calcium salt, magnesium salt, etc.), ammonium salt, an inorganicacid salt (e.g. hydrochloride, hydrobromide, sulfate, phosphate,carbonate, bicarbonate, etc.), a salt with or organic base or acid, forexample, an amine salt (e.g. trimethylamine salt, triethylamine salt,pyridine salt, procaine salt, picoline salt, dicyclohexylamine salt,N,N'-dibenzylethylenediamine salt, N-methylglucamine salt,diethanolamine salt, triethanolamine salt,tris(hydroxymethylamino)methane salt, phenethylbenzylamine salt, etc.),an organic carboxylic or sulfonic acid salt (e.g. acetate, maleate,lactate, tartrate, mesylate, benzenesulfonate, tosylate, etc.), a basicor acidic amino acid salt (e.g. arginine salt, aspartic acid salt,glutamic acid salt, lysine salt, serine salt, etc.) and the like.

The processes for preparing the object compounds (I) of the presentinvention are explained in details in the following.

Process A: N-Acylation

A compound (I) or its salt can be prepared by reacting a compound (II)or its reactive derivative at the amino or a salt thereof with acompound (III) or its reactive derivative at the carboxy or a saltthereof according to a conventional manner of so-called amidationreaction well known in β-lactam chemistry.

Suitable reactive derivative at the amino group of the compound (II) mayinclude a conventional reactive derivative as used in a wide variety ofamidation reaction, for example, isocyanato, isothiocyanato, aderivative formed by the reaction of a compound (II) with a silylcompound (e.g. trimethylsilylacetamide, bis(trimethylsilyl)acetamide,etc.), with an aldehyde compound (e.g. acetaldehyde, isopentaldehyde,benzaldehyde, salicylaldehyde, phenylacetaldehyde, p-nitrobenzaldehyde,m-chlorobenzaldehyde, p-chlorobenzaldehyde, hydroxynaphthoaldehyde,furfural, thiophenecarboaldehyde, etc., or the corresponding hydrate,acetal, hemiacetal or enolate thereof), with a ketone compound (e.g.acetone, methyl ethyl ketone, methyl isobutyl ketone, acetylacetone,ethyl acetoacetate, etc., or the corresponding ketal, hemiketal orenolate thereof), with phosphorus compound (e.g., phosphorusoxychloride, phosphorus chloride, etc.), or with a sulfur compound (e.g.thionyl chloride, etc.), and the like.

Suitable salt of the compound (II) may be referred to those asexemplified for the compound (I).

Suitable reactive derivative at the carboxy group of the compound (III)may include, for example, an acid halide, an acid anhydride, anactivated amide, an activated ester, and the like, and preferably acidhalide such as acid chloride, acid bromide; a mixed acid anhydride withan acid such as substituted phosphoric acid (e.g. dialkylphosphoricacid, phenylphosphoric acid, diphenylphosphoric acid, dibenzylphosphoricacid, halogenated phosphoric acid, etc.), dialkylphosphorous acid,sulfurous acid, thiosulfuric acid, sulfuric acid, alkylcarbonic acid,aliphatic carboxylic acid (e.g. pivalic acid, pentanoic acid,isopentanoic acid, 2-ethylbutyric acid, trichloroacetic acid, etc.),aromatic carboxylic acid (e.g. benzoic acid, etc.); a symmetrical acidhydride; an activated acid amide with imidazole, 4-substitutedimidazole, dimethylpyrazole, triazole or tetrazole; an activated ester(e.g. cyanomethyl ester, methoxymethyl ester, dimethylaminomethyl ester,vinyl ester, propargyl ester, p-nitrophenyl ester, 2,4-dinitrophenylester, trichlorophenyl ester, pentachlorphenyl ester, mesylphenyl ester,phenylazophenyl ester, phenyl thioester, p-nitrophenyl thioester,p-cresyl thioester, carboxymethyl thioester, pyranyl ester, pyridylester, piperidyl ester, 8-quinolyl thioester, an ester with a N-hydroxycompound such as N,N-dimethylhydroxylamine, 1-hydroxy-2-(1H)-pyridone,N-hydroxysuccinimide, N-hydroxyphthalimide, 1-hydroxybenzotriazole,1-hydroxy-6-chlorobenzotriazole, etc.), and the like.

Suitable salt of the compound (III) may include a salt with an inorganicbase such as alkali metal salt (e.g. sodium salt, potassium salt, etc.)and an alkaline earth metal salt (e.g. calcium salt, magnesium salt,etc.) a salt with an organic base such as tertiary amine (e.g.trimethylamine salt, triethylamine salt, N,N-dimethylaniline salt,pyridine salt, etc.), a salt with an inorganic acid (e.g. hydrochloride,hydrobromide, etc.) and the like.

The suitable reactive derivatives of the compounds (II) and (III) canoptionally be selected from the above according to the kind of thecompounds (II) and (III) to be used practically, and to the reactionconditions.

The reaction is usually carried out in a conventional solvent such aswater, acetone, dioxane, acetonitrile, chloroform, benzene, methylenechloride, ethylene chloride, tetrahydrofuran, ethyl acetate,N,N-dimethylformamide, pyridine or any other solvent which does notadversely influence the reaction, or an optional mixture thereof.

When the acylating agent (III) is used in a form of free acid or salt inthis reaction, the reaction is preferably carried out in the presence ofa condensing agent such as a carbodiimide compound (e.g.N,N'-dicyclohexylcarbodiimide,N-cyclohexyl-N'-morpholinoethylcarbodiimide,N-cyclohexyl-N'-(4-diethylaminocyclohexyl)carbodiimide,N,N'-diethylcarbodiimide, N,N'-diisopropylcarbodiimide,N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide, etc.), a bisimidazolidecompound (e.g. N,N'-carbonylbis(2-methylimidazole), etc.), an iminecompound (e.g. pentamethyleneketene-N-cyclohexylimine,diphenylketene-N-cyclohexylimine, etc.), an olefinic or acetylenic ethercompound (e.g. ethoxyacetylene, β-chlorovinylethyl ether, etc.),1-(4-chlorobenzenesulfonyloxy)-6-chloro-1H-benzotriazole,N-ethylbenzisoxazolium salt, N-ethyl-5-phenylisoxazolium-3'-sulfonate, aphosphorus compound (e.g. polyphosphoric acid, trialkyl phosphite, ethylpolyphosphate, isopropyl polyphosphate, phosphorus oxychloride,phosphorus trichloride, diethylchlorophosphite, orthophenylenechlorophosphite, etc.), thionyl chloride, oxalyl chloride, Vilsmeierreagent prepared by the reaction of the dimethylformamide with thionylchloride, phosphoryl chloride, phosgene or the like.

The reaction temperature is not critical, and the reaction is usuallycarried out under cooling or at ambient temperature.

In order to obtain a syn isomer of the compound (I) selectively and inhigh yield, it is preferable to use a syn isomer of the acylating agent(III), and to conduct the reaction under a selected reaction condition.That is, a syn isomer of the compound (I) can be obtained selectivelyand in high yield by conducting the reaction of a compound (II) with asyn isomer of the acylating agent (III), for example, in the presence ofa Vilsmeier reagent as mentioned above and under around neutralcondition.

The object compound (I) and its salt are useful as an antimicrobialagent, and a part thereof can also be used as a starting material in theother processes as explained below.

Process B: Thioetherification

A compound (I) or its salt can be prepared by reacting a compound (IV)or its salt with a thiol compound (V) or its salt.

"Acyl" group for R⁵ of the starting compound (IV) may be lower alkanoylhaving 2 to 6 carbon atom(s) (e.g. acetyl, propionyl, etc.), aroyl (e.g.benzoyl, toluoyl, etc.) or the like.

Suitable salt of the compound (IV) may be referred to those asexemplified for the compound (I).

Suitable salt of the thiol compound (V) may include a metal salt such asalkali metal salt (e.g. sodium salt, potassium salt, etc.), and acidsalt such as hydrohalide (e.g. hydrochloride, hydrobromide, etc.) or thelike.

This reaction may be carried out in a solvent such as water, acetone,chloroform, nitrobenzene, methylene chloride, ethylene chloride,dimethylformamide, methanol, ethanol, diethyl ether, tetrahydrofuran,dimethylsulfoxide, buffer solution or any other solvent which does notadversely affect the reaction, preferably in ones having strongpolarities. Among the solvents, hydrophilic solvents may be used in amixture with water. The reaction is preferably conducted in weekly basicor around neutral condition. When the compound (IV) and/or the thiolcompound (V) is used in a free form, the reaction is preferablyconducted in the presence of a base, for example, inorganic base such asalkali metal hydroxide, alkali metal carbonate, alkali metalbicarbonate, organic base such as trialkylamine, pyridine, and the like.The reaction temperature is not critical, and the reaction is usuallycarried out at ambient temperature or under warming.

Process C: Elimination of amino-protective group in R_(a) ¹

A compound (I_(c)) or its salt can be prepared by subjecting a compound(I_(c')) or its salt to elimination reaction of the protective group inthe protected amino for R_(a) ¹.

The elimination reaction may be conducted in accordance with aconventional method such as hydrolysis, reduction or the like. Thesemethods may be selected according to the kind of the protective group tobe eliminated.

The hydrolysis may include a method using an acid (acidic hydrolysis), abase (basic hydrolysis) or hydrazine, and the like. Among these methods,acidic hydrolysis is one of the common and preferable methods foreliminating the protective group such as an acyl lower alkanoyl (e.g.formyl, acetyl, etc.), aralkyl (e.g. trityl, etc.), lower alkoxycarbonyl (e.g. tert-pentyloxycarbonyl, etc.), substituted loweralkanoyl, substituted lower alkoxycarbonyl, substituted or unsubstitutedar(lower)alkoxycarbonyl, lower cycloalkoxycarbonyl, substitutedphenylthio, substituted alkylidene, substituted aralkylidene,substituted cycloalkylidene or the like. Suitable acid to be used inthis acidic hydrolysis may include an organic or inorganic acid such asformic acid, trifluoroacetic acid, benzenesulfonic acid,p-toluenesulfonic acid, hydrochloric acid, cation-exchange resin, andthe like. Preferable acid is the one which can easily be separated outfrom the reaction product by a conventional manner such asneutralization or distillation under reduced pressure, for example,formic acid, trifluoroacetic acid, hydrochloric acid or the like. Theacid suitable for the reaction can be selected in consideration of thechemical property of the starting compound and the product as well asthe kind of the protective group to be eliminated. The acidic hydrolysiscan be conducted in the presence or absence of a solvent. Suitablesolvent may be a conventional organic solvent, water or a mixturethereof which does not adversely influence this reaction. The hydrolysisusing trifluoroacetic acid is accelerated by addition of anisole.

The basic hydrolysis can be preferably, applied for eliminating theprotective group such as an acyl group, for example, haloalkanoyl (e.g.trifluoroacetyl, etc.) and the like. Suitable base may include, forexample, an inorganic base such as alkali metal hydroxide (e.g. sodiumhydroxide, potassium hydroxide, etc.), alkaline earth metal hydroxide(e.g. magnesium hydroxide, calcium hydroxide, etc.), alkali metalcarbonate (e.g. sodium carbonate, potassium carbonate, etc.), alkalineearth metal carbonate (e.g. magnesium carbonate, calcium carbonate,etc.), alkali metal bicarbonate (e.g. sodium bicarbonate, potassiumbicarbonate, etc.), alkaline earth metal phosphate (e.g. magnesiumphosphate, calcium phosphate, etc.), alkali metal hydrogen phosphate(e.g. disodium hydrogen phosphate, dipotassium hydrogen phosphate,etc.), or the like, and an organic base such as alkali metal acetate(e.g. sodium acetate, potassium acetate, etc.), trialkylamine (e.g.trimethylamine, triethylamine, etc.), picoline, N-methylpyrrolidine,N-methylmorpholine, 1,5-diazobicyclo[4,3,0]-5-nonene,1,4-diazabicyclo[2,2,2]-octane, 1,5-diazabicyclo[5,4,0]-7-undecene,anion-exchange resin or the like. The hydrolysis using a base is oftencarried out in water or a conventional organic solvent or a mixturethereof.

The hydrolysis using hydrazine can be applied for eliminating theprotective group such as dibasic acyl, for example, succinyl, phthaloylor the like.

The reduction can be applied for eliminating the protective group suchas acyl, for example, halo(lower)-alkoxycarbonyl (e.g.trichloroethoxycarbonyl, etc.), substituted or unsubstitutedar(lower)alkoxycarbonyl (e.g. benzyloxycarbonyl,p-nitrobenzyloxycarbonyl, etc.), 2-pyridylmethoxycarbonyl, etc.,ar(lower)alkyl (e.g. benzyl, benzhydryl, trityl, etc.) and the like.Suitable reduction may include, for example, reduction using an alkalimetal borohydride (e.g. sodium borohydride, etc.), conventionalcatalytic hydrogenolysis and the like.

And further, the protective group such as halo(lower)alkoxycarbonyl or8-quinolyloxycarbonyl can be eliminated by treatment with a heavy metalsuch as copper, zinc or the like.

The reaction temperature is not critical and may be optionally selectedin consideration of the chemical property of the starting compound andreaction product as well as the kind of the N-protective group and themethod to be applied, and the reaction is preferably carried out under amild condition such as under cooling, at ambient temperature or slightlyelevated temperature.

The process includes in its scope the cases that the amino-protectivegroups of the protected amino(lower)alkyl moiety in R² and R³ areeliminated and/or that the protected carboxy for R⁴ is simultaneouslytransformed into the free carboxy group in the course of the abovereaction or in the post-treatment.

As to this process, it is to be understood that the purpose of thisprocess lies in providing the generally more active compound (I_(c)) byeliminating the protective group in the protected amino group of thecompound (I_(c')) prepared by the other processes as mentioned above orbelow.

Process D: Elimination of amino-protective group in R_(a) ³

A compound (I_(d)) or its salt can be prepared by subjecting a compound(I_(d')) or its salt to elimination reaction of the protective group inthe protected amino(lower)alkyl moiety for R_(a) ³.

The elimination reaction can be conducted in the same manner as theabove Process C.

This process includes in its scope the cases that the amino-protectivegroups of the protected amino for R¹ and protected amino(lower)alkyl forR² are eliminated and/or that the protected carboxy for R⁴ issimultaneously transformed into the free carboxy group in the course ofthe above reaction or the post-treatment.

Process E: Elimination of hydroxy-protective group

A compound (I_(e)) or its salt can be prepared by subjecting a compound(I_(e')), or its salt to elimination reaction of the protective group ofthe protected hydroxy(lower)alkyl for R_(a) ².

This reaction may be conducted in a similar manner to that of theaforementioned Process C.

This process includes in its scope the cases that the amino-protectivegroup of the protected amino for R¹ and/or the protectedamino(lower)alkyl moiety in R² and R³ are eliminated in the course ofthe reaction or post-treatment.

The compound obtained in accordance with the processes as explainedabove can be isolated and purified in a conventional manner.

In case that the object compound (I) has free amino for R¹, freeamino(lower)alkyl-substituted heterocyclic group for R³ and/or freecarboxy for R⁴, it may be transformed into its pharmaceuticallyacceptable salt by a conventional method.

Process F: Elimination of amino-protective group in R_(c) ²

A compound (I_(f)) or its salt can be prepared by subjecting a compound(I_(f')) or its salt to elimination reaction of the protective group inthe protected amino(lower)alkyl for R_(c) ².

The elimination reaction can be conducted in the same manner as theabove Process C.

This process includes in its scope, the cases that amino-protectivegroup of the protected amino for R¹ and/or the protected amino moiety inR³ is eliminated, and/or the protected carboxy for R⁴ is simultaneouslytransformed into the free carboxy group in the course of the abovereaction or the post-treatment.

Process G: Carboxy Formation in R_(e) ²

A compound (I_(g)) of its salt can be prepared by subjecting a compound(I_(g')) or its salt to a reaction of transforming an esterified carboxymoiety into carboxy moiety.

The reaction is carried out by conventional method, such as hydrolysis,reduction or the like.

The hydrolysis can be conducted in the same manner as the above ProcessC. The reduction may be conducted with a conventional reducing agentwhich is used for transforming the esterified carboxy group to a freecarboxy group, for example, an alkali metal borohydride (e.g., sodiumborohydride etc.), palladium carbon, palladium oxide, platinum oxide,and the like.

The reaction temperature is not critical and may be suitable selected inaccordance with the kind of the ester and the method to be applied, andthe present reaction is preferably carried out under a mild conditionsuch as under cooling, at ambient temperature or slightly elevatedtemperature.

Process H: Carboxy formation

A compound (I_(h)) or its salt can be prepared by subjecting a compound(I_(h')) or its salt to elimination reaction of the protective group atthe carboxy for R_(a) ⁴.

The elimination reaction of this process can be conducted in the samemanner as the above Process G.

The compound obtained in accordance with the processes as explainedabove can be isolated and purified in a conventional manner.

In case that the object compound (I) has free amino for R¹, freeamino(lower)alkyl for R², free amino(lower)alkyl-substitutedheterocyclic group for R³ and/or free carboxy for R⁴, it may betransformed into its pharmaceutically acceptable salt by a conventionalmethod.

The object compound (I) and its pharmaceutically acceptable salt exhibithigh antimicrobial activities inhibiting the growth of a wide variety ofpathogenic microorganisms including Gram-positive and Gram-negativebacteria and are useful as antimicrobial agents.

In order to show the utility of the compound (I) the test data of somerepresentative compounds (I) are shown in the following.

1. In Vitro Antibacterial Activity (1) Test Method

In vitro antibacterial activity was determined by the two-foldagar-plate dilution method as described below.

One loopful of an overnight culture of each test strain in Trypticasesoy broth was streaked on heart infusion agar (HI-agar) containinggraded concentration of the test compound and incubated at 37° C. for 20hours. The minimal inhibitory concentration (MIC) was expressed inμg/ml.

(2) Test Compound

    ______________________________________    No.1      7-[2-(2-aminothiazol-4-yl)-2-(3-amino-              propoxyimino)acetamido]-3-(5-aminomethyl-              1,3,4-thiadiazol-2-yl)thiomethyl-3-cephem-              4-carboxylic acid trihydrochloride (syn              isomer)    No.2      7-[2-(2-aminothiazol-4-yl)-2-(2-amino-              ethoxyimino)acetamido]-3-[1-(2-aminoethyl)-              1H-tetrazol-5-yl]thiomethyl-3-cephem-4-              carboxylic acid trihydrochloride (syn              isomer)    No.3      7-[2-(2-aminothiazol-4-yl)-2-carboxymethoxy-              iminoacetamido]-3-[1-(2-aminoethyl)-1H-              tetrazol-5-ylthiomethyl]-3-cephem-4-carbo-              xylic acid (syn isomer)    No.4      7-[2-(2-aminothiazol-4-yl)-2-(2-hydroxy-              ethoxyimino)acetamido]-3-[1-(2-hydroxy-              ethyl)-1H-tetrazol-5-ylthiomethyl]-3-              cephem-4-carboxylic acid hydrochloride              (syn isomer)    No.5      7-[2-(2-aminothiazol-4-yl)-2-propargyloxy-              iminoacetamido]-3-[1-(2-hydroxyethyl)-1H-              tetrazol-5-ylthiomethyl]-3-cephem-4-              carboxylic acid (syn isomer)    ______________________________________

(3) Test Results

MIC (μg/ml)

    ______________________________________            Compound No.    Test strains              1        2       3      4      5    ______________________________________    Escherichia              0.200    0.390   0.100  0.200  0.390    coli NIHJ JC-2    Klebsiella              0.200    0.100   0.100  0.050  0.100    pneumoniae 20    Proteus   1.560    1.560   0.050  0.050  0.050    vulgaris 2    Serratia  100.000  3.130   12.500 25.000 50.000    marcescens 35    Enterobacter              6.250    1.560   12.500 25.000 50.000    eloacae 60    Enterobacter              6.250    0.390   6.250  12.500 6.250    aerogenes 20    ______________________________________

For prophylactic and/or therapeutic administration, the compound (I) ofthe present invention is used in the form of conventional pharmaceuticalpreparation which contains said compound, as an active ingredient, inadmixture with pharmaceutically acceptable carriers such as an organicor inorganic solid or liquid excipient which is suitable for oral,parenteral or external administration. The pharmaceutical preparationsmay be in solid form such as capsule, tablet, dragee, ointment orsuppository, or in liquid form such as solution, suspension, oremulsion. If needed, there may be included in the above preparationsauxiliary substances, stabilizing agents, wetting or emulsifying agents,buffers and the other commonly used additives.

While the dosage of the compounds may vary from and also depend upon theage and conditions of the patient, a kind of disease and a degree of theinfection, and further a kind of the active compound (I) to be applied,etc., an average single dose of about 50 mg., 100 mg., 250 mg. and 500mg. of the active compound (I) is sufficient for treating infectiousdiseases caused by pathogenic bacteria. In general, the active compound(I) can be administered in an amount between 1 mg/kg and 100 mg/kg,preferably 5 mg/kg and 50 mg/kg.

Following preparations and examples are given only for explanation ofthis invention in more detail.

Preparation 1

(1) Allyl bromide (2.91 g) was added dropwise to a stirred suspension ofethyl 2-(2-tritylaminothiazol-4-yl)-2-hydroxyiminoacetate (syn isomer,10 g), N,N-dimethylformamide (100 ml) and potassium carbonate (4.54 g)under ice cooling over 5 minutes, and stirred at the same temperaturefor 4 hours. After adding water (200 ml) to the resultant solution, thesolution was extracted with diethyl ether twice. The extract was washedwith a saturated aqueous solution of sodium chloride and dried overmagnesium sulfate. The solution was concentrated in vacuo, and theresidue was triturated with a solution of n-hexane and diethyl ether.The precipitates were collected by filtration to give ethyl2-(2-tritylaminothiazol-4-yl)-2-allyloxyimioacetate (syn isomer, 9.4 g),mp. 130° to 132° C.

I.R. ν_(max) ^(Nujol) : 3380, 1735, 1520, 1550 cm⁻¹

N.M.R. δ (DMSO-d₆, ppm): 1.08 (3H, t, J=7 Hz), 3.96 (2H, q, J=7 Hz),4.54 (2H, broad d, J=5 Hz), 5.05-5.5 (2H, m), 5.6-6.3 (1H, m), 6.90(15H, broad s), 7.74 (1H, s)

(2) A solution of ethyl2-(2-tritylaminothiazol-4-yl)-2-allyloxyiminoacetate (syn isomer, 8.7g), 50% formic acid (42.5 ml) and tetrahydrofuran (42.5 ml) was stirredat 60° C. for 40 minutes. After concentrating the resultant solution invacuo, the residue was dissolved in ethyl acetate, washed with anaqueous solution of sodium bicarbonate and a saturated aqueous solutionof sodium chloride in turn, and dried over magnesium sulfate. Afterconcentrating the resultant solution in vacuo, the residue was subjectedto column chromatography on silica gel with benzene and ethyl acetate inturn, to give ethyl 2-(2-aminothiazol-4-yl)-2-allyloxyiminoacetate (synisomer, 3.7 g), mp. 102° to 104° C.

I.R. ν_(max) ^(Nujol) : 3460, 3260, 3130, 1725, 1620, 1540, 1460 cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 1.25 (3H, t, J=7 Hz), 4.30 (2H, q, J=7 Hz), 4.61(2H, d,d, J=5 Hz), 5.0-5.5 (2H, m), 5.6-6.5 (1H, m), 6.95 (1H, s), 7.28(2H, s)

(3) A solution of ethyl 2-(2-aminothiazol-4-yl)-2-allyloxyiminoacetate(syn isomer, 3.6 g), 2 N aqueous solution of sodium hydroxide (14.1 ml),tetrahydrofuran (14.1 ml) and methanol (15 ml) was stirred at 40° C. for1.5 hours. The resultant solution was concentrated in vacuo, and theresidue was dissolved in water. After the solution was adjusted to pH2.8 with 10% hydrochloric acid under ice cooling, the precipitates werecollected by filtration, washed with water and acetone in turn and driedto give 2-(2-aminothiazol-4-yl)-2-allyloxyiminoacetic acid (syn isomer,1.91 g), mp. 187° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3350, 1630, 1580, 1460 cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 4.61 (2H, d, J=6 Hz), 5.1-5.5 (2H, m), 5.7-6.2(1H, m), 6.84 (1H, s), 7.25 (2H, broad s)

Preparation 2

(1) A mixture of ethyl 2-hydroxyimino-3-oxobutyrate (syn isomer, 56.7g), N,N-dimethylformamide (280 ml), potassium carbonate (72.3 g) andpropargyl bromide (43 g) was stirred at room temperature for 4 hrs. Thereaction mixture was treated in a conventional manner to give ethyl2-propargyloxyimino-3-oxobutyrate (syn isomer, 71.2 g).

I.R. ν_(max) ^(Film) : 3280, 3220, 2120, 1735, 1670 cm⁻¹

(2) A mixture of ethyl 2-propargyloxyimino-3-oxobutyrate (syn isomer,71.2 g), acetic acid (81 ml) and sulfuryl chloride (50.2 g) was stirredat 40° C. for 10 minutes and then at room temperature for 5.5 hrs. Thereaction mixture was treated in a conventional manner to give ethyl4-chloro-3-oxo-2-propargyloxyiminobutyrate (syn isomer, 61.6 g.), oil.

I.R. ν_(max) ^(Film) : 3300, 2130, 1745, 1720, 1675 cm⁻¹

N.M.R. δ(CCl₄, ppm): 1.39 (3H, t, J=7 Hz), 2.57 (1H, t, J=2 Hz), 4.36(2H, q, J=7 Hz), 4.56 (2H, s), 4.86 (2H, d, J=2 Hz)

(3) A mixture of ethyl 4-chloro-4-oxo-2-propargyloxyiminobutyrate (synisomer, 61 g), thiourea (20 g), sodium acetate 3-hydrate (35.8 g), water(150 ml), and ethanol (180 ml) was stirred at 40° C. for 1.25 hrs. Thereaction mixture was treated in a conventional manner to give ethyl2-(2-aminothiazol-4-yl)-2-propargyloxyiminoacetate (syn isomer, 35.6 g).

I.R. ν_(max) ^(Nujol) : 3290, 2220, 1729 cm⁻¹

N.M.R. δ (DMSO-d₆, ppm): 1.28 (3H, t, J=7 Hz), 3.49 (1H, t, J=3 Hz),4.31 (2H, q, J=7 Hz), 4.76 (2H, d, J=3 Hz), 6.95 (1H, s), 7.29 (2H, s).

(4) A mixture of ethyl2-(2-aminothiazol-4-yl)-2-propargyloxyiminoacetate (syn isomer, 2.8 g.),methanol (23 ml.), tetrahydrofuran (20 ml.) and 1 N aqueous solution ofsodium hydroxide (22.17 ml.) was stirred at 30° C. for 5 hrs. Thereaction mixture was treated in a conventional manner to give2-(2-aminothiazol-4-yl)-2-propargyloxyiminoacetic acid (syn isomer,1.924 g.).

I.R. Ξ_(max) ^(Nujol) : 2190, 1740 cm⁻¹

N.M.R. δ (DMSO-d₆, ppm): 3.47 (1H, t, J=1.5 Hz), 4.74 (2H, d, J=1.5 Hz),6.90 (1H, s)

Preparation 3

(1) A mixture of ethyl 2-hydroxyimino-3-oxobutyrate (syn isomer, 15.7g.), 2-bromoethyl benzoate (27.5 g.), potassium carbonate (20.7 g.),N,N-dimethylformamide (25 ml.) and ethyl acetate (25 ml.) was stirred atroom temperature for 4 hrs. The reaction mixture was treated in aconventional manner to give ethyl2-(2-benzoyloxyethoxyimino)-3-oxobutyrate (syn isomer, 28 g.).

(2) A solution of ethyl 2-(2-benzoyloxyethoxyimino)-3-oxobutyrate (synisomer, 28 g.), sulfuryl chloride (13.5 g.) and acetic acid (30 ml.) wasstirred at 40° C. for 10 minutes and at room temperature for 5.5 hrs.The reaction mixture was treated in a conventional manner to give ethyl2-(2-benzoyloxyethoxyimino)-4-chloro-3-oxobutyrate (syn isomer, 29 g.).

(3) A mixture of ethyl2-(2-benzoyloxyethoxyimino)-4-chloro-3-oxobutyrate (syn isomer, 29 g.),thiourea (7.76 g.), sodium acetate (8.37 g.), water (75 ml.) and ethanolwas stirred at 40° C. for an hour. The reaction mixture was treated in aconventional manner to give ethyl2-(2-aminothiazol-4-yl)-2-(2-benzoyloxyethoxyimino)acetate (syn isomer,9 g.).

N.M.R. δ (DMSO-d₆, ppm): 1.28 (3H, t, J=7 Hz), 4.34 (2H, q, J=7 Hz),4.56 (4H, m), 6.44 (2H, broad s), 6.68 (1H, s), 7.68-7.34 (3H, m), 8.06(2H, d,d, J=8 Hz, 2 Hz)

(4) A solution of ethyl2-(2-aminothiazol-4-yl)-2-(2-benzoyloxyethoxyimino)acetate (syn isomer,8.5 g.) in a mixture of 1 N aqueous sodium hydroxide (35 ml.), methanol(40 ml.) and tetrahydrofuran (40 ml.) was stirred at 40° C. for 9 hrs.and at room temperature for 12 hrs. The reaction mixture was treated ina conventional manner to give2-(2-aminothiazol-4-yl)-2-(2-hydroxyethoxyimino)acetic acid (syn isomer,3.3 g.).

I.R. ν_(max) ^(Nujol:) 3350, 3075, 1680, 1620 cm⁻¹

N.M.R. δ (DMSO-d₆, ppm): 3.64 (2H, t, J=5 Hz), 4.10 (2H, t, J=5 Hz),6.84 (1H, s), 7.16 (2H, m)

(5) A solution of formic acid (1.6 g.) and acetic anhydride (3.6 g.) wasstirred at 50° C. for an hour. After cooling,2-(2-aminothiazol-4-yl)-2-(2-hydroxyethoxyimino)acetic acid (syn isomer,1 g.) was added to the solution and stirred at room temperature for 3hours. Diisopropyl ether was added to the resultant solution, and theprecipitates were filtered out. The filtrate was concentrated in vacuo,and the residue was pulverized with diisopropyl ether. The precipitateswere collected by filtration to give2-(2-formamidothiazol-4-yl)-2-(2-formyloxyethoxyimino)-acetic acid (synisomer, 0.7 g.).

I.R. ν_(max) ^(Nujol) : 3200, 1710, 1690 cm⁻¹

N.M.R. δ (DMSO-d₆, ppm): 4.38 (4H, s), 7.58 (1H, s), 8.26 (1H, s), 8.54(1H, s)

Preparation 4

(1) A mixture of chloromethylthiomethane (7.97 g.), powdered potassiumiodide (15.1 g.) and acetone (79 ml.) was stirred at room temperaturefor an hour, the resulting mixture was filtered and washed with a smallamount of acetone. The washings and the filtrate were combined and addedto a stirred suspension of ethyl2-(2-formamidothiazol-4-yl)-2-hydroxyiminoacetate (syn isomer, 17.5 g.)and powdered potassium carbonate (15.5 g.) in acetone (300 ml.). Themixture was stirred at room temperature for 3 hours, filtered and washedwith acetone. The washings and the filtrate were combined andconcentrated in vacuo. The residue was dissolved in ethyl acetate,washed with a saturated aqueous solution of sodium chloride twice, driedover magnesium sulfate and concentrated in vacuo. The oily residue wassubjected to column chromatography on silica gel and eluted withchloroform to give ethyl2-(2-formamidothiazol-4-yl)-2-methylthiomethoxyiminoacetate (syn isomer,2.4 g.), mp. 130° to 131° C.

I.R. ν_(max) ^(Nujol) : 3160, 3125, 3050, 1740, 1695 cm⁻¹

N.M.R. δ (DMSO-d₆, ppm): 1.32 (3H, t, J=7 Hz), 2.22 (3H, s), 4.38 (2H,q, J=7 Hz), 5.33 (2H, s), 7.67 (1H, s), 8.56 (1H, s)

(2) A mixture of ethyl2-(2-formamidothiazol-4-yl)-2-methylthiomethoxyiminoacetate (syn isomer,2.4 g.), 1 N aqueous sodium hydroxide (23.8 ml.) and methanol (19.8 ml.)was stirred at 30° C. for 2.5 hours. The resultant solution was adjustedto pH 7 with 10% hydrochloric acid and methanol was distilled off invacuo. The aqueous solution was adjusted to pH 1 with 10% hydrochloricacid under ice cooling, and extracted with ethyl acetate three times.The extracts were washed with a saturated aqueous solution of sodiumchloride, dried over magnesium sulfate and concentrated in vacuo to give2-(2-formamidothiazol-4-yl)-2-methylthiomethoxyiminoacetic acid (synisomer, 1.13 g.), mp. 157° C. (dec.).

I.R. ν_(max) ^(Nujol) : 3210, 3160, 3075, 1700, 1555 cm⁻¹

N.M.R. δ (DMSO-d₆, ppm): 2.24 (3H, s), 5.31 (2H, s), 7.61 (1H, s), 8.57(1H, s), 12.73 (1H, s)

Preparation 5

Hydrazine hydrate (13.2 g.) was added to a suspension ofN-phthalimidoxyethylphthalimide (38.4 g.) in ethanol (400 ml.) at 65° C.and stirred at 65° to 70° C. for an hour. After adding conc.hydrochloric acid (28 ml.) and water (280 ml.) to the resultant solutionunder ice-cooling, the insoluble substance was filtered off. Thefiltrate was concentrated in vacuo, washed with ethyl acetate andadjusted to pH 7.0 with 10% sodium hydroxide solution. After washing thesolution with ethyl acetate, ethanol (400 ml.) and2-(2-formamidothiazol-4-yl)glyoxylic acid (16.0 g.) were added to thesolution and stirred at room temperature at pH 4.5 to 4.8 for 2 hours.The solvent was removed in vacuo, and ethyl acetate was added to theresidue. The solution was adjusted to pH 0.3 with conc. hydrochloricacid. The aqueous solution was separated and adjusted to pH 5.6 with 10%sodium hydroxide solution. To the solution were added dioxane (600 ml.),triethylamine (16.0 g.) and2-tert-butoxycarbonyloxyimino-2-phenylacetonitrile (23.6 g.), andstirred at room temperature overnight. After removing the solvent invacuo, the residue was washed with ethyl acetate. The aqueous solutionwas separated and ethyl acetate was added thereto. The mixture wasadjusted to pH 2.0 with 10% hydrochloric acid under ice-cooling. Theorganic layer was separated and washed with saturated aqueous sodiumchloride solution. The solution was dried over magnesium sulfate andevaporated in vacuo. The residue was pulverized with diisopropyl etherto give2-(2-formamidothiazol-4-yl)-2-(2-tert-butoxycarbonylaminoethoxyimino)aceticacid (syn isomer, 13.3 g.).

I.R. ν_(max) ^(Nujol) : 3140, 1698, 1604 cm⁻¹

N.M.R. δ (DMSO-d₆, ppm): 1.37 (9H, s), 3.20 (2H, m), 3.97 (2H, m), 7.33(1H, s), 8.50 (1H, s)

Preparation 6

Trimethylsilylphosphite (8.0 g.) was added to a solution of2-(2-formamidothiazol-4-yl)-2-(2-azidoethoxyimino)acetic acid (synisomer, 3.4 g.) and bis(trimethylsilyl)acetamide (5.4 g.) in pyridine(20 ml.) and stirred at room temperature for 20 hours. Water (10 ml.)was added to the resultant solution at 5° to 10° C. and evaporated invacuo. Water and ethyl acetate were added to the residue and adjusted topH 1.5 with 10% hydrochloric acid. The aqueous layer was separated andadjusted to pH 7.0 with 10% sodium hydroxide. Dioxane (30 ml.),triethylamine (6.0 g.) and2-tert-butoxycarbonyloxyimino-2-phenylacetonitrile (4.9 g.) were addedto the resultant solution, and stirred at room temperature for 15 hours.After removing the solvent in vacuo, ethyl acetate was added to theresidue and adjusted to pH 6.0 with 10% hydrochloric acid underice-cooling. The aqueous solution was separated and ethyl acetate wasadded thereto. The mixture was adjusted to pH 2.0 with 10% hydrochloricacid under ice-cooling. The ethyl acetate layer was separated, washedwith saturated sodium chloride solution and dried over magnesiumsulfate. The solution was concentrated in vacuo and the residue waspulverized with diisopropyl ether to give2-(2-formamidothiazol-4-yl)-2-(2-tert-butoxycarbonylaminoethoxyimino)aceticacid (syn isomer, 1.7 g.).

I.R. ν_(max) ^(Nujol) : 3140, 1698, 1604 cm⁻¹

Preparation 7

100% Hydrazine hydrate (10.0 g.), N-phthaliminoxypropylphthalimide (35g.), 2-(2-formamidothiazol-4-yl)-glyoxylic acid (8.86 g.) and2-tert-butoxycarbonyloxyimino-2-phenylacetonitrile (29.6 g.) weretreated in a similar manner to that of Example 5 to give2-(2-formamidothiazol-4-yl)-2-(3-tert-butoxycarbonylaminopropoxyimino)aceticacid (syn isomer, 6.0 g.).

I.R. ν_(max) ^(Nujol) : 3360, 3170, 1700, 1570, 1540 cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 1.40 (9H, s), 1.80 (2H, m), 3.07 (2H, m), 4.18(2H, t, J=7 Hz), 7.57 (1H, s), 8.57 (1H, s), 12.7 (1H, broad s)

Preparation 8

(1) A solution of N-(3-aminopropyl)acetamide (146 g) in dioxane (710 ml)was added to a solution of 97% sodium hydroxide (52 g) in water (620 ml)and then carbon disulfide (96 g) was added dropwise thereto over 35minutes at -1° to 3° C. The mixture was stirred for 1 hour at 0° to 2°C. To the mixture containing sodium N-(3-acetamidopropyl)dithiocarbamatewas added dropwise methyl iodide (179 g) over 35 minutes at 0° to 5° C.and then the resulting mixture was stirred for 3 hours at the sametemperature. Dioxane was distilled off in vacuo from the reactionmixture and the residue was extracted with ethyl acetate (300 ml, 200ml×4). The extracts were dried over magnesium sulfate and concentratedin vacuo to give oil of methyl N-(3-acetamidopropyl)dithiocarbamate(193.18 g).

(2) A mixture of a solution of methylN-(3-acetamidopropyl)dithiocarbamate (193 g) in dioxane (610 ml) and asolution of sodium azide (79.42 g) in water (500 ml) was refluxed understirring for 4 hours. Dioxane was distilled off and the remainingaqueous layer was washed with diethyl ether (150 ml×2), adjusted to pH 1with 17.5% hydrochloric acid, and cooled in an ice bath. Precipitateswere collected by filtration and washed with ice-water to give whitepowder of 1-(3-acetamidopropyl)-1H-tetrazol-5-thiol (91.75 g), mp. 152°to 154° C.

N.M.R. (d₆ -DMSO) δ: 1.87 (3H, s), 1.97 (2H, m), 3.17 (2H, m), 4.28 (2H,t, J=7 Hz), 7.9 (1H, broad s), 15.0 (1H, broad s)

(3) A mixture of 1-(3-acetamidopropyl)-1H-tetrazole-5-thiol (85 g) and 6N hydrochloric acid (1 l) was refluxed for 75 minutes under stirring.The reaction mixture was concentrated in vacuo and precipitates werecollected by filtration and washed with hexane and diethyl ether to give1-(3-aminopropyl)-1H-tetrazole-5-thiol hydrochloride (67.15 g).

N.M.R. (D₂ O) δ: 2.45 (2H, m), 3.23 (2H, t, J=7 Hz), 4.50 (2H, t, J=7Hz)

(4) A solution of 2-t-butoxycarbonyloxyimino-2-phenylacetonitrile (12.3g) in dioxane (30 ml) was added under ice-cooling to a stirred solutionof 1-(3-aminopropyl)-1H-tetrazole-5-thiol hydrochloride (9.78 g) andtriethylamine (11.1 g) in a mixture of dioxane (25 ml) and water (25ml), and then the resulting mixture was stirred for 1.75 hours atambient temperature. Dioxane was distilled off and to the residue wereadded diethyl ether and a small amount of water. After shaking, theaqueous layer was separated and the organic layer was extracted twicewith 10% potassium carbonate. The extracts combined with the separatedaqueous layer were washed three times with diethyl ether, adjusted to pH1 with hydrochloric acid and extracted with diethyl ether. The extractwas washed with water, dried and evaporated in vacuo. The residual oil(10.92 g) was pulverized with diisopropyl ether to give1-[3-(N-t-butoxycarbonylamino)propyl]-1H-tetrazole-5-thiol (9.6 g), mp.75° to 77° C.

I.R. (Nujol): 3380, 3260, 1650, 1530, 1170, 1050 cm⁻¹

N.M.R. (CDCl₃) δ: 1.50 (9H, s), 2.14 (2H, m), 3.25 (2H, m), 4.39 (2H, t,J=7 Hz), 4.9-6.7 (1H, broad)

EXAMPLE 1

(1) A solution of 2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetic acid(syn isomer, 1.4 g) in dry ethyl acetate (20 ml) was added to a mixtureof dry dimethylformamide (0.5 g), dry ethyl acetate (2.0 ml) andphosphoryl chloride (1.0 g) to give an activated acid solution. On theother hand,7-amino-3-[1-(2-hydroxyethyl)-1H-tetrazol-5-yl-thiomethyl]-3-cephem-4-carboxylicacid (2.0 g) and trimethylsilylacetamide (5.9 g) were added to dry ethylacetate (40 ml), stirred at 40° C. and cooled to -10° C. To the solutionwas added the activated acid solution at -5° to -10° C. and stirred atthe same temperature for an hour. Water (40 ml) was added to thereaction mixture and adjusted to pH 7.0 with sodium bicarbonate. Theaqueous layer was separated and washed with ethyl acetate and diethylether successively. After removing the remaining ethyl acetate anddiethyl ether by bubbling with nitrogen gas, the aqueous solution wasadjusted to pH 2.0 with conc. hydrochloric acid and stirred for 30minutes. The precipitates were collected by filtration, washed withchilled water and then dried over phosphorus pentoxide to give7-[2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-[1-(2-hydroxyethyl-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (syn isomer, 2.10 g).

I.R. (Nujol) νmax: 1760, 1660 cm⁻¹

N.M.R. (DMSO-d₆) δppm: 3.46-4.04 (4H, m), 3.90 (3H, s), 4.12-4.53 (4H,m), 5.12 (1H, d, J=5.0 Hz), 5.79 (1H, d,d, J=5.0 Hz, 8.0 Hz), 7.42 (1H,s), 8.52 (1H, s), 9.67 (1H, d, J=8.0 Hz)

(2) A mixture of7-[2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-[1-(2-hydroxyethyl-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (syn isomer, 2.0 g), conc.hydrochloric acid (0.73 g) and methanol(14.0 ml) was stirred at room temperature for 3 hrs. and evaporated invacuo. The residue was dissolved in aqueous solution of sodiumbicarbonate and then acidified to pH 3 with 10% hydrochloric acid. Theprecipitates were collected, washed with chilled water and dried overphosphorus pentoxide to give7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-[1-(2-hydroxyethyl-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (syn isomer, 1.1 g)

I.R. (Nujol) νmax: 3350, 1775, 1670, 1635 cm⁻¹

N.M.R. (DMSO-d₆) δppm: 3.33-4.10 (4H, m), 3.83 (3H, s), 4.10-4.61 (4H,m), 5.11 (1H, d, J=4.3 Hz), 5.77 (1H, d,d, J=4.3 Hz, 8.0 Hz), 6.76 (1H,s), 9.60 (1H, d, J=8.0 Hz)

EXAMPLE 2

(1) A solution of7-amino-3-[1-(2-hydroxyethyl-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (2.0 g) and trimethylsilylacetamide (5.9 g) in dry ethyl acetate(40.0 ml) and a solution of2-(2-formamidothiazol-4-yl)-2-allyloxyiminoacetic acid (syn isomer, 1.6g), dry dimethylformamide (0.5 g) and phosphoryl chloride (1.0 g) in dryethyl acetate 22.0 ml were treated in a similar manner to that ofExample 1-(1) to give7-[2-(2-formamidothiazol-4-yl)-2-allyloxyiminoacetamido]-3-[1-(2-hydroxyethyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (syn isomer, 2.79 g). PG,50

I.R. (Nujol) νmax: 3180, 1770, 1665 cm⁻¹

N.M.R. (DMSO-d₆) δppm: 3.48-3.95 (4H, m), 4.03-4.50 (4H, m), 4.50-4.78(2H, m), 5.01-5.54 (3H, m), 5.65-6.60 (2H, m), 7.41 (1H, s), 8.53 (1H,s), 9.67 (1H, d, J=8.5 Hz)

(2)7-[2-(2-Formamidothiazol-4-yl)-2-allyloxyiminoacetamido]-3-[1-(2-hydroxyethyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (syn isomer, 2.7 g) was treated with conc.hydrochloric acid (0.94g) in a similar manner to that of Example 1-(2) to give7-[2-(2-aminothiazol-4-yl)-2-allyloxyiminoacetamido]-3-[1-(2-hydroxyethyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid hydrochloride (syn isomer, 2.72 g).

I.R. (Nujol) νmax: 3340, 3210, 1772, 1730, 1665 cm⁻¹

N.M.R. (DMSO-d₆) δppm: 3.44-3.83 (4H, m), 4.00-4.40 (4H, m), 4.57 (2H,m), 4.95-5.47 (3H, m), 5.53-6.74 (4H, m), 6.82 (1H, s), 9.77 (1H, d,J=8.0 Hz)

EXAMPLE 3

(1) A solution of7-amino-3-[1-(2-hydroxyethyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (2.0 g) and trimethylsilylacetamide (5.9 g) in dry ethyl acetate(40.0 ml) and a solution of2-(2-formamidothiazol-4-yl)-2-propargyloxyiminoacetic acid (syn isomer,1.6 g), dry dimethylformamide (0.5 g) and phosphoryl chloride (1.0 g) indry ethyl acetate (52.0 ml) were treated in a similar manner to that ofExample 1-(1) to give7-[2-(2-formamidothiazol-4-yl)-2-propargyloxyiminoacetamido]-3-[1-(2-hydroxyethyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (syn isomer, 2.00 g).

I.R. (Nujol) νmax: 3260, 1780, 1670 cm⁻¹

N.M.R. (DMSO-d₆) δppm: 3.49 (1H, m), 3.57-4.05 (4H, m), 4.09-4.67 (4H,m), 4.79 (2H, m), 5.15 (1H, d, J=5.0 Hz), 5.81 (1H, d,d, J=5.0 Hz, 8.0Hz), 7.46 (1H, s), 8.55 (1H, s), 9.76 (1H, d, J=8.0 Hz)

(2)7-[2-(2-Formamidothiazol-4-yl)-2-propargyloxyiminoacetamido]-3-[1-(2-hydroxyethyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (syn isomer, 1.9 g) was treated with conc.hydrochloric acid (0.67g) in a similar manner to that of Example 1-(2) to give7-[2-(2-aminothiazol-4-yl)-2-propargyloxyiminoacetamido]-3-[1-(2-hydroxyethyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (syn isomer, 1.19 g).

I.R. (Nujol) νmax: 3300, 1775, 1670 cm⁻¹

N.M.R. (DMSO-d₆) δppm: 3.47 (1H, m), 3.56-4.00 (4H, m), 4.00-4.56 (4H,m), 4.72 (2H, m), 5.14 (1H, d, J=5.0 Hz), 5.79 (1H, d,d, J=5.0 Hz, 8.0Hz), 6.82 (1H, s), 9.67 (1H, d, J=8.0 Hz).

EXAMPLE 4

(1) A solution of7-amino-3-[1-(2-hydroxyethyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (2.16 g) in 50% aqueous acetone (22 ml) and a solution of2-(2-formamidothiazol-4-yl)-2-methylthiomethoximinoacetic acid (synisomer, 1.5 g), dimethylformamide (0.48 g) and phosphoryl chloride (1.01g) in tetrahydrofuran (15 ml) were treated in a similar manner to thatof Example 1-(1) to give7-[2-(2-formamidothiazol-4-yl)-2-methylthiomethoxyiminoacetamido]-3-[1-(2-hydroxyethyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (syn isomer, 2.58 g).

I.R. (Nujol) νmax: 3420, 3250, 3050, 1770, 1660, 1540 cm⁻¹

N.M.R. (DMSO-d₆) δppm: 2.23 (3H, s), 3.52-3.97 (4H, m), 4.11-4.62 (4H,m), 5.17 (1H, d, J=5 Hz), 5.28 (2H, s), 5.85 (1H, d,d, J=5 Hz, 8 Hz),7.47 (1H, s), 8.54 (1H, s), 9.75 (1H, d, J=8 Hz), 12.69 (1H, broad s)

(2)7-[2-(2-Formamidothiazol-4-yl)-2-methylthiomethoxyiminoacetamido]-3-[1-(2-hydroxyethyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (syn isomer, 2.5 g) was treated with conc.hydrochloric acid (0.88g) in a similar manner to that of Example 1-(2) to give7-[2-(2-aminothiazol-4-yl)-2-methylthiomethoxyiminoacetamido-]-3-[1-(2-hydroxyethyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (syn isomer, 1.8 g).

I.R. (Nujol) νmax: 3350, 1780, 1670, 1630, 1540 cm⁻¹

N.M.R. (DMSO-d₆) δppm: 2.16 (3H, s), 3.3-3.9 (4H, m), 4.0-4.6 (4H, m),5.08 (1H, d, J=5 Hz), 5.15 (2H, s), 5.76 (1H, d, d, J=5 Hz, 8 Hz), 6.76(1H, s), 7.22 (2H, broad s), 9.77 (1H, d, J=8 Hz)

EXAMPLE 5

(1) A solution of7-amino-3-[1-(2-hydroxyethyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (2.0 g) and trimethylsilylacetamide (5.9 g) in dry ethyl acetate(40 ml) and a solution of2-(2-formamidothiazol-4-yl)-2-(2-formyloxyethoxyimino)acetic acid (synisomer, 1.8 g), dry dimethylformamide (0.5 g) and phosphoryl chloride(1.0 g) in dry ethyl acetate (22.0 ml) were treated in a similar mannerto that of Example 1-(1) to give7-[2-(2-formamidothiazol-4-yl)-2-(2-formyloxyethoxyimino)acetamido]-3-[1-(2-hydroxyethyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (syn isomer, 1.10 g).

I.R. (Nujol) νmax: 3170, 1770, 1660 cm⁻¹

N.M.R. (DMSO-d₆) δppm: 3.43-3.88 (4H, m), 3.95-4.60 (8H, m), 5.11 (1H,d, J=5.0 Hz), 5.79 (1H, d,d, J=5.0 Hz, 8.0 Hz), 7.42 (1H, s), 8.22 (1H,s), 8.51 (1H, s), 9.65 (1H, d, J=8.0 Hz)

(2)7-[2-(2-Formamidothiazol-4-yl)-2-(2-formyloxyethoxyiminoacetamido]-3-[1-(2-hydroxyethyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (syn isomer, 1.0 g) was treated with conc. hydrochloric acid (0.66g) in a similar manner to that of Example 1-(2) to give7-[2-(2-aminothiazol-4-yl)-2-(2-hydroxyethoxyimino)acetamido]-3-[1-(2-hydroxyethyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (syn isomer, 0.97 g).

I.R. (KBr) νmax: 3300, 3100, 2970, 1770, 1735, 1640 cm⁻¹

N.M.R. (DMSO-d₆) δppm: 3.50-3.95 (6H, m), 4.00-4.63 (6H, m), 5.14 (1H,d, J=5.0 Hz), 5.28-6.85 (3H, m), 6.93 (1H, s) 9.71 (1H, d, J=8.0 Hz)

EXAMPLE 6

(1) Vilsmeier reagent was prepared from N,N-dimethylformamide (0.65 g.)and phosphoryl chloride (1.4 g.) in a usual manner.2-(2-Formamidothiazol-4-yl)-2-(3-tert-butoxycarbonylaminopropoxyimino)aceticacid (syn isomer, 3.0 g.) was added to a stirred suspension of theVilsmeier reagent in ethyl acetate (30 ml.) under ice-cooling andstirred at the same temperature for 30 minutes [Solution A].Trimethylsilylacetamide (8.1 g.) was added to a stirred suspension of7-amino-3-(5-tert-butoxycarbonylaminomethyl-1,3,4-thiadiazol-2-yl)thiomethyl-3-cephem-4-carboxylicacid (3.7 g.) in ethyl acetate (40 ml.) and stirred at room temperaturefor 30 minutes. The solution A was added to the solution all at once at-30° C., and stirred at -10° to -40° C. for an hour. Water and ethylacetate (100 ml.) were added to the resultant mixture at -10° C., andthe ethyl acetate layer was separated. Water (100 ml.) was added to theethyl acetate layer and adjusted to pH 7.0 with saturated aqueous sodiumbicarbonate solution. The aqueous layer was separated. Ethyl acetate wasadded to the solution and adjusted to pH 3.8 with 10% hydrochloric acidunder ice-cooling. The ethyl acetate layer was separated and washed withsaturated sodium chloride solution. The solution was dried overmagneisum sulfate and concentrated in vacuo. The residue was pulverizedwith diisopropyl ether to give7-[2-(2-formamidothiazol-4-yl)-2-(3-tert-butoxycarbonylaminopropoxyimino)acetamido]-3-(5-tert-butoxycarbonylaminomethyl-1,3,4-thiadiazol-2-yl)-thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 1.6 g).

I.R. ν_(max) ^(Nujol) : 3400-3200, 1780, 1690, 1530 cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 1.42 (9H, s), 1.80 (2H, m), 3.70 (2H, m), 3.73(2H, broad s), 4.00-4.87 (6H, m), 5.20 (1H, d, J=5 Hz), 5.87 (1H, dd,J=5 Hz, 8 Hz), 7.10 (1H, s), 8.55 (1H, s), 9.67 (1H, d, J=8 Hz)

(2) A mixture of7-[2-(2-formamidothiazol-4-yl)-2-(3-tert-butoxycarbonylaminopropoxyimino)acetamido]-3-(5-tert-butoxycarbonylaminomethyl-1,3,4-thiadiazol-2-yl)-thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 1.5 g.), conc. hydrochloric acid (0.8 ml.), methanol(30 ml.) and tetrahydrofuran (30 ml.) was stirred at room temperaturefor 3 hours. After evaporation, methanol was added to the residue. Thesolution was evaporated in vacuo again, and the residue was dissolved inwater (30 ml.). The solution was adjusted to pH 3.5 with saturatedaqueous sodium bicarbonate solution under ice-cooling. The solution wassubjected to column chromatography on macroporous non-ionic adsorptionresin "Diaion HP-20" (trademark: Mitsubishi Chemical Industries Ltd.)and eluted with 30% aqueous isopropyl alcohol. The eluate wasconcentrated in vacuo and lyophilized to give7-[2-(2-aminothiazol-4-yl)-2-(3-aminopropoxyimino)acetamido]-3-(5-aminomethyl-1,3,4-thiadiazol-2-yl)thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 0.6 g.).

I.R. ν_(max) ^(Nujol) : 3400-3100, 1770, 1600, 1610, 1530 cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 2.03 (2H, m), 3.00 (2H, m), 3.70 (2H, m),3.93-4.83 (6H, m), 5.13 (1H, d, J=5 Hz), 5.77 (1H, dd, J=5 Hz, 8 Hz),6.78 (1H, s), 9.62 (1H, d, J=8 Hz)

EXAMPLE 7

(1)2-(2-Formamidothiazol-4-yl)-2-(3-tert-butoxycarbonylaminopropoxyimino)aceticacid (syn isomer, 2.6 g.), N,N-dimethylformamide (0.6 g.), phosphorylchloride (1.2 g.),7-amino-3-[1-(2-tert-butoxycarbonylaminoethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid (3.2 g.), trimethylsilylacetamide (5.5 g.) and ethyl acetate (65ml.) were treated in a similar manner to that of Example 6-(1) to give7-[2-(2-formamidothiazol-4-yl)-2-(3-tert-butoxycarbonylaminopropoxyimino)acetamido]-3-[1-(2-tert-butoxycarbonylaminoethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 1.3 g.).

I.R. ν_(max) ^(Nujol) : 3350, 3200, 1780, 1690, 1650, 1540 cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 1.40 (18H, s), 1.83 (2H, m), 3.08 (2H, m),3.20-3.66 (2H, m), 3.77 (2H, broad s), 3.92-4.73 (6H, m), 5.20 (1H, d,J=5 Hz), 5.88 (1H, dd, J=5 Hz, 8 Hz), 7.45 (1H, s), 8.58 (1H, s), 9.67(1H, d, J=8 Hz), 12.72 (1H, broad s)

(2)7-[2-(2-Formamidothiazol-4-yl)-2-(3-tert-butoxycarbonylaminopropoxyimino)acetamido]-3-[1-(2-tertbutoxycarbonylaminoethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 2.8 g.), conc. hydrochloric acid (1.6 ml.) andmethanol (60 ml.) were treated in a similar manner to that of Example6-(2) to give7-[2-(2-aminothiazol-4-yl)-2-(3-aminopropoxyimino)acetamido]-3-[1-(2-aminoethyl)-1H-tetrazol-5-yl]-thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 1.1 g).

I.R. ν_(max) ^(Nujol) : 3500-3100, 1770, 1660, 1640-1560, 1540 cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 2.00 (2H, m), 2.97 (2H, m), 3.17-4.17 (4H, m),4.17-4.93 (6H, m), 5.08 (1H, d, J=5 Hz), 5.75 (1H, m), 6.77 (1H, s),9.55 (1H, m)

EXAMPLE 8

(1)2-(2-Formamidothiazol-4-yl)-2-(2-tert-butoxycarbonylaminoethoxyimino)aceticacid (syn isomer, 2 g.), N,N-dimethylformamide (0.45 g.), phosphorylchloride (1.03 g.), 7-amino-3-[1-(2-tert-butoxycarbonylaminoethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylic acid (2.6 g.),trimethylsilylacetamide (5.9 g.) and ethyl acetate (50 ml.) were treatedin a similar manner to that of Example 6-(1) to give7-[2-(2-formamidothiazol-4-yl)-2-(2-tert-butoxycarbonylaminoethoxyimino)acetamido]-3-[1-(2-tert-butoxycarbonylaminoethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 3.7 g.).

I.R. ν_(max) ^(Nujol) : 3300, 1780, 1680, 1540 cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 1.37 (18H, s), 3.07-3.60 (4H, m), 3.75 (2H,broad s), 3.93-4.57 (4H, m), 5.20 (1H, d, J=5Hs), 5.90 (1H, dd, J=8 Hz,5Hz), 7.47 (1H, s), 8.57 (1H, s), 9.62 (1H, d, J=8 Hz), 12.72 (1H, broads)

(2) Conc. hydrochloric acid (3.2 g.) was added to a solution of7-[2-(2-formamidothiazol-4-yl)-2-(2-tert-butoxycarbonylaminoethoxyimino)acetamido]-3-[1-(2-tert-butoxycarbonylaminoethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 3.5 g.) in methanol (50 ml.) and stirred at roomtemperature for 2 hours. After removing the solvent in vacuo, methanolwas added to the residue. The mixture was concentrated in vacuo again.The precipitates were collected by filtration and washed with diethylether to give7-[2-(2-aminothiazol-4-yl)-2-(2-aminoethoxyimino)acetamido]-3-[(2-aminoethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid trihydrochloride (syn isomer, 2.8 g.).

I.R. ν_(max) ^(Nujol) : 3500-3100, 1770, 1700, 1670, 1620, 1560, 1540cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 3.0-3.53 (4H, m), 3.80 (2H, m), 4.17-4.83 (6H,m), 5.20 (1H, d, J=5 Hz), 5.80 (1H, dd, J=8 Hz, 5Hz), 7.10 (1H, s), 9.93(1H, d, J=8 Hz)

EXAMPLE 9

(1)2-(2-Formamidothiazol-4-yl)-2-(2-tert-butoxycarbonylaminoethoxyimino)aceticacid (syn isomer, 2 g.), N,N-dimethylformamide (0.45 g.), phosphorylchloride (1.03 g.),7-amino-3-(5-tert-butoxycarbonylaminomethyl-1,3,4-thiadiazol-2-yl)thiomethyl-3-cephem-4-carboxylicacid (2.6 g.), trimethylsilylacetamide (5.9 g.) andbis(trimethylsilyl)acetamide (3.4 g.) and ethyl acetate (50 ml.) weretreated in a similar manner to that of Example 6-(1) to give7-[2-(2-formamidothiazol-4-yl)-2-(2-tert-butoxycarbonylaminoethoxyimino)acetamido]-3-(5-tert-butoxycarbonylaminomethyl-1,3,4-thiadiazol-2-yl)thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 4.0 g.).

I.R. ν_(max) ^(Nujol) : 3400, 3200, 1775, 1680, 1535 cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 3.12-3.43 (2H, m), 3.60 (2H, m), 4.03 (2H, m),4.37 (2H, q, J=13 Hz), 4.17-4.57 (2H, m), 5.08 (1H, d, J=5 Hz), 5.57(1H, dd, J=8 Hz, 5 Hz), 7.30 (1H, s), 8.40 (1H, s), 9.43 (1H, d, J=8Hz), 12.55 (1H, broad s)

(2)7-[2-(2-Formamidothiazol-4-yl)-2-(2-tert-butoxycarbonylaminoethoxyimino)acetamido]-3-(5-tert-butoxycarbonylaminomethyl-1,3,4-thiadiazol-2-yl)thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 3.7 g.), conc. hydrochloric acid (2.9 g.) and methanol(50 ml.) were treated in a similar manner to that of Example 8-(2) togive7-[2-(2-aminothiazol-4-yl)-2-(2-aminoethoxyimino)acetamido]-3-(5-aminomethyl-1,3,4-thiadiazol-2-yl)thiomethyl-3-cephem-4-carboxylicacid trihydrochloride (syn isomer, 2.9 g.).

I.R. ν_(max) ^(Nujol) : 3500-3100, 1770, 1700, 1670, 1625, 1570, 1540cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 3.30 (2H, m), 3.73 (2H, m), 4.10-4.80 (6H, m),5.17 (1H, d, J=5 Hz), 5.77 (1H, dd, J=8 Hz, 5 Hz), 7.0 (1H, s), 9.90(1H, d, J=8 Hz)

EXAMPLE 10

(1) Vilsmeier reagent was prepared from N,N-dimethylformamide (0.4 g.)and phosphoryl chloride (0.8 g.) in dry ethyl acetate (1.6 g.). Dryethyl acetate (16 ml.) and2-(2-formamidothiazol-4-yl)-2-(3-tert-butoxycarbonylaminopropoxyiminoaceticacid (syn isomer, 1.6 g.) were added to the Vilsmeier reagent [SolutionA]. The solution A was added dropwise to a solution of7-amino-3-(4-amino-5-methyl-4H-1,2,4-triazol-3-yl)-thiomethyl-3-cephem-4-carboxylicacid (1.5 g.), and sodium bicarbonate (1.1 g.) in water (9 ml.) andacetone (9 ml.) at -2° to 3° C. while adjusting to pH 7.0 to 8.0 withtriethylamine, and stirred at the same temperature for 30 minutes. Ethylacetate and water were added to the resultant solution. The aqueouslayer was separated and washed with ethyl acetate. The solution wasconcentrated in vacuo and the residue was adjusted to pH 2.5 withphosphoric acid under ice-cooling. The precipitates were collected byfiltration, washed with water and dried over magnesium sulfate to give7-[2-(2-formamidothiazol-4-yl)-2-(3-tert-butoxycarbonylaminopropoxyimino)acetamido]-3-(4-amino-5-methyl-4H-1,2,4-triazol-3-yl)thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 1.55 g.).

I.R. ν_(max) ^(Nujol) : 1770, 1675 cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 1.39 (9H, s), 1.80 (2H, m), 2.31 (3H, s), 3.01(2H, m), 3.67 (2H, m), 4.19 (4H, m), 5.13 (1H, d, J=4.0 Hz), 5.78 (1H,dd, J=4.0 Hz, 8.0 Hz), 7.41 (1H, s), 8.53 (1H, s), 9.62 (1H, d, J=8 Hz)

(2) A solution of7-[2-(2-formamidothiazol-4-yl)-2-(3-tert-butoxycarbonylaminopropoxyimino)acetamido]-3-(4-amino-5-methyl-4H-1,2,4-triazol-3-yl)thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 1.45 g.), and conc. hydrochloric acid (1.1 g.) inmethanol (10.2 ml.) was treated in a similar manner to that of Example8-(2) to give7-[2-(2-aminothiazol-4-yl)-2-(3-aminopropoxyimino)acetamido]-3-(4-amino-5-methyl-4H-1,2,4-triazol-3-yl)thiomethyl-3-cephem-4-carboxylicacid trihydrochloride (syn isomer, 1.21 g.)

I.R. ν_(max) ^(Nujol) : 1780, 1670, 1630 cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 2.11 (2H, m), 2.90 (2H, m), 3.79 (2H, m),4.04-4.75 (4H, m), 5.23 (1H, d, J=5.0 Hz), 5.82 (1H, dd, J=5.0 Hz, 9.0Hz), 7.01 (1H, s), 9.94 (1H, d, J=9.0 Hz)

EXAMPLE 11

(1) 2-(2-Formamidothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoaceticacid (syn isomer, 1.62 g.) was added to a solution ofN,N-dimethylformamide (432 mg.) and phosphoryl chloride (905 mg.) intetrahydrofuran (16 ml.) and treated in a similar manner to that ofExample 6-(1). The solution was added to a solution of7-amino-3-[1-(2-tert-butoxycarbonylaminoethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid (3.0 g.) and triethylamine in 50% aqueous acetone (30 ml.) at -5°to -3° C. and pH 7 to 7.5 and stirred for 30 minutes. Ethyl acetate wasadded to the resultant solution and adjusted to pH 2.0 with 10%hydrochloric acid. After removing the insoluble substance by filtration,water and ethyl acetate were added to the filtrate. The ethyl acetatelayer was separated and washed with saturated sodium chloride solution.The solution was dried over magnesium sulfate and concentrated in vacuo.The residue was pulverized with diethyl ether and the precipitates werecollected by filtration to give7-[2-(2-formamidothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-[1-(2-tert-butoxycarbonylaminoethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 2.93 g.).

I.R. ν_(max) ^(Nujol) : 3270, 1790, 1695, 1550, 1460 cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 1.28 (9H, s), 1.38 (9H, s), 3.27 (2H, broad s),3.63 (2H, s), 4.24 (4H, s), 4.50 (2H, s), 5.04 (1H, d, J=5 Hz), 5.72(1H, dd, J=5 Hz), 8 Hz), 8.36 (1H, s), 9.42 (1H, d, J=8 Hz), 12.52 (1H,s)

(2) A solution of7-[2-(2-formamidothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-[1-(2-tert-butoxycarbonylaminoethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 3.5 g.) and conc. hydrochloric acid (1.9 g.) inmethanol (35 ml.) was stirred at room temperature for 1.5 hours. Afterconcentration, the residue was pulverized with diethyl ether to give7-[2-(2-aminothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-[1-(2-tert-butoxycarbonylaminoethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid hydrochloride (syn isomer, 3.32 g.).

I.R. ν_(max) ^(Nujol) : 3350 (broad), 1775, 1720, 1680, 1635, 1570, 1550cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 1.42 (9H, s), 1.57 (9H, s), 3.36 (2H, broad s),3.72 (2H, q, J=18 Hz), 4.32 (2H, s), 4.64 (4H, broad s), 5.15 (1H, d,J=5 Hz), 5.74 (1H, dd, J=5 Hz, 8 Hz), 7.02 (1H, s), 7.36 (2H, broad s),9.75 (1H, d, J=8 Hz)

(3) Trifluoroacetic acid (12.8 ml.) was added portionwise to a chilledsuspension of7-[2-(2-aminothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-[1-(2-tert-butoxycarbonylaminoethyl)-1H-tetrazol-5-yl]-thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 3.2 g.) in anisole (3.2 ml.), and stirred at roomtemperature for 70 minutes. After removing the solvent from theresultant mixture in vacuo, the residue was triturated with diethylether. The precipitates were collected by filtration, dried anddissolved in water (40 ml.). The solution was adjusted to pH 4.8 with10% sodium hydroxide solution under ice-cooling. The solution wassubjected to column chromatography on macroporous non-ionic adsorptionresin "Diaion HP-20" (trademark: Mitsubishi Chemical Industries Ltd.)and eluted with 20% aqueous isopropyl alcohol. The eluate wasconcentrated in vacuo and lyophilized to give7-[2-(2-aminothiazol-4-yl)-2-carboxymethoxyiminoacetamido]-3-[1-(2-aminoethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 1.2 g.).

I.R. ν_(max) ^(Nujol) : 3300, 3170, 1760, 1660, 1530 cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 3.44 (2H, broad s), 3.72 (2H, s), 3.9-5.2 (7H,m), 5.08 (1H, d, J=5 Hz), 5.73 (1H, broad s), 6.87 (1H, s), 7.22 (2H,broad s)

EXAMPLE 12

(1) 2-(2-Formamidothiazol-4-yl)-2-ethoxycarbonylmethoxyiminoacetic acid(syn isomer, 1.32 g.), N,N-dimethylformamide (447 mg.), phosphorylchloride (939 mg.),7-amino-3-[1-(2-tert-butoxycarbonylaminoethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid (3.0 g.), tetrahydrofuran (13 ml.) and 50% aqueous acetone (30 ml.)were treated in a similar manner to that of Example 11-(1) to give7-[2-(2-formamidothiazol-4-yl)-2-ethoxycarbonylmethoxyiminoacetamido]-3-[1-(2-tert-butoxycarbonylaminoethyl)-1H-tetrazol-5-yl]-thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 2.78 g.).

I.R. ν_(max) ^(Nujol) : 3260 (broad), 1780, 1690 (shoulder), 1540 cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 1.21 (3H, t, J=7 Hz), 3.37 (2H, broad s), 3.72(2H, broad s), 4.16 (2H, q, J=7 Hz), 4.34 (4H, s), 4.73 (2H, s), 5.16(1H, d, J=5 Hz), 5.85 (1H, dd, J=5 Hz, 8 Hz), 7.48 (1H, s), 8.58 (1H,s), 9.52 (1H, d, J=8 Hz), 12.24 (1H, s)

(2)7-[2-(2-formamidothiazol-4-yl)-2-ethoxycarbonylmethoxyiminoacetamido]-3-[1-(2-tert-butoxycarbonylaminoethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 2.7 g.), conc. hydrochloric acid (1.52 g.) and ethanol(27 ml.) were treated in a similar manner to that of Example 11-(2) togive(7-[2-(2-aminothiazol-4-yl)-2-ethoxycarbonylmethoxyiminoacetamido]-3-[1-(2-aminoethyl)-1H-tetrazol-5-yl]-thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 0.57 g.).

I.R. ν_(max) ^(Nujol) : 3300, 3170, 1760, 1670, 1530 cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 1.22 (3H, t, J=7 Hz), 3.42 (2H, broad s), 3.60(2H, broad s), 4.16 (2H, q, J=7 Hz), 4.42 (2H, broad s), 5.07 (1H, d,J=5 Hz), 5.72 (1H, broad s), 6.80 (1H, s), 7.25 (2H, broad s), 9.48 (1H,broad s)

EXAMPLE 13

(1)2-(2-Formamidothiazol-4-yl)-2-(2-tert-butoxycarbonylaminoethoxyimino)aceticacid (syn isomer, 2 g.), N,N-dimethylformamide (0.45 g.), phosphorylchloride (1.0 g.),7-amino-3-[1-(2-hydroxyethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid (2 g.), trimethylsilylacetamide (5.9 g.) and ethyl acetate (50 ml.)were treated in a similar manner to that of Example 6-(1) to give7-[2-(2-formamidothiazol-4-yl)-2-(2-tert-butoxycarbonylaminoethoxyimino)acetamido]-3-[1-(2-hydroxyethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 2.7 g.).

I.R. ν_(max) ^(Nujol) : 3300, 1780, 1680, 1540 cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 1.35 (9H, s), 3.33 (2H, m), 3.75 (4H, m), 4.07(2H, m), 4.33 (4H, m), 5.17 (1H, d, J=5 Hz), 5.87 (1H, dd, J=8 Hz, 5Hz), 7.40 (1H, s), 8.50 (1H, s), 9.53 (1H, d, J=8 Hz), 12.57 (1H, m)

(2)7-[2-(2-Formamidothiazol-4-yl)-2-(2-tert-butoxycarbonylaminoethoxyimino)acetamido]-3-[1-(2-hydroxyethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 2.5 g.), conc. hydrochloric acid (1.9 g.) and methanol(40 ml.) were treated in a similar manner to that of Example 6-(2) togive7-[2-(2-aminothiazol-4-yl)-2-(2-aminoethoxyimino)acetamido]-3-[1-(2-hydroxyethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 1.4 g.).

I.R. ν_(max) ^(Nujol) : 3300, 3170, 1765, 1660, 1600, 1530 cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 3.13 (2H, m), 3.50 (2H, m), 3.73 (2H, m),4.60-3.93 (6H, m), 4.97 (1H, d, J=5 Hz), 5.67 (1H, dd, J=8 Hz, 5 Hz),6.75 (1H, s), 9.43 (1H, m)

EXAMPLE 14

(1) Vilsmeier reagent was prepared from N-N-dimethylformamide (0.53 g)and phosphoryl chloride (1.1 g) in a usual manner.2-(2-Formamidothiazol-4-yl)-2-propargyloxyiminoacetic acid (syn isomer,1.5 g) was added to a suspension of the Vilsmeier reagent in dry ethylacetate (12 ml) under ice-cooling and stirred at the same temperaturefor 30 minutes [Solution A].

Trimethylsilylacetamide (4.72 g) was added to a stirred suspension of7-amino-3-[1-(2-tert-butoxycarbonylaminoethyl)-1-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid (3.05 g) in ethyl acetate (30 mg) and the mixture was stirred atroom temperature for 30 minutes. To the solution was added the above[Solution A] all at once at -30° C. and stirred at -10° to -40° C. foran hour. Water and ethyl acetate were added to the reaction mixture at-10° C.

The ethyl acetate layer was separated and extracted with an aqueoussolution of sodium bicarbonate (pH 7.0). To the aqueous solution wasadded ethyl acetate and adjusted to pH 5.0 under ice-cooling. The ethylacetate layer was separated, washed with a saturated aqueous solution ofsodium chloride, dried over magnesium sulfate and concentrated in vacuoto give7-[2-(2-formamidothiazol-4-yl)-2-propargyloxyiminoacetamido]-3-[1-(2-tert-butoxycarbonylaminoethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 1.16 g).

I.R. ν_(max) ^(Nujol) : 3275, 1780, 1690, 1540 cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 1.32 (9H, s), 3.03-3.58 (3H, m), 3.7 (2H, broads), 4.18-4.5 (4H, m), 4.73 (2H, m), 5.13 (1H, d, J=5 Hz), 5.82 (1H, dd,J=5 Hz, 8 Hz), 7.43 (1H, s), 8.53 (1H, s), 9.78 (1H, d, J=8 Hz)

(2) A mixture of7-[2-(2-formamidothiazol-4-yl)-2-propargyloxyiminoacetamido]-3-[1-(2-tert-butoxycarboxylaminoethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 0.63 g) and conc. hydrochloric acid (0.3 g) inmethanol (5 ml) was stirred at 35° C. for 1.5 hours.

After evaporating the solvent from the resultant mixture, in vacuo, theresidue was dissolved in methanol (10 ml). Methanol was added to theresidue and evaporated in vacuo again. The residue was pulverized withdiisopropyl ether to give7-[2-(2-aminothiazol-4-yl)-2-propargyloxyiminoacetamido]-3-[1-(2-aminoethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid dihydrochloride (syn isomer, 0.52 g).

I.R. ν_(max) ^(Nujol) : 3300-3100, 1770, 1700(sh), 1670, 1630,1540^(cm-1)

N.M.R. δ(DMSO-d₆, ppm): 3.35 (2H, m), 3.55 (1H, m), 3.75 (2H, broad s),5.05-4.15 (4H, m), 4.78 (2H, m), 5.13 (1H, d, J=5 Hz), 5.73 (1H, dd, J=5Hz, 8 Hz), 6.93 (1H, s), 9.78 (1H, d, J=8 Hz)

EXAMPLE 15

(1) Vilsmeier reagent was prepared from N,N-dimethylformamide (0.6 g)and phosphoryl chloride (1.3 g) in dry ethyl acetate (2.4 ml) in a usualmanner. Dry ethyl acetate (18 ml) and2-(2-formamidothiazol-4-yl)-2-allyloxyiminoacetic acid (syn isomer, 1.8g) were added to the solution of Vilsmeier reagent subsequently, andthen stirred [Solution A]. Trimethylsilylacetamide (6.1 g) was added toa stirred suspension of7-amino-3-[1-(2-tert-butoxycarbonylaminoethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid (3.0 g) in dry ethyl acetate (60 ml) and stirred at 40° C. for 30minutes.

The solution A was added to the stirred solution at -5° to -10° C. andstirred at the same temperature for 30 minutes. After adding water tothe resultant mixture, the insoluble substance was collected byfiltration and dissolved in tetrahydrofuran. The ethyl acetate layer wasseparated from the filtrate and combined with the tetrahydrofuransolution. The solution was washed with a saturated sodium chloridesolution and dried over magnesium sulfate. After concentrating thesolution in vacuo, the residue was pulverized with diisopropyl ether,collected by filtration and washed with diisopropyl ether to give7-[2-(2-formamidothiazol-4-yl)-2-allyloxyiminoacetamido]-3-[1-(2-tert-butoxycarbonylaminoethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 2.98 g).

I.R. ν_(max) ^(Nujol) : 1780, 1670^(cm-1)

N.M.R. δ(DMSO-d₆, ppm): 1.35 (9H, s), 3.37 (2H, m), 3.67 (2H, m), 4.37(4H, m), 4.66 (2H, m), 5.03-5.62 (3H, m), 5.67-6.34 (2H, m), 7.43 (1H,s), 8.54 (1H, s), 9.71 (1H, d, J=8.0 Hz)

(2) A suspension of7-[2-(2-formamidothiazol-4-yl)-2-allyl-oxyiminoacetamido]-3-[1-(2-tert-butoxycarbonylaminoethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylicacid (syn isomer, 2.9 g) and conc. hydrochloric acid (1.7 g) in methanol(20.3 ml) was stirred at room temperature for 2.7 hours. After removingthe solvent in vacuo, methanol (20 ml) was added to the residue andevaporated in vacuo again. Water and ethyl acetate were added to theresidue and adjusted to pH 7.5 with sodium bicarbonate. The aqueoussolution was separated, washed with ethyl acetate and the remainingorganic solvent was removed in vacuo. The aqueous solution was adjustedto pH 3.7 with 10% hydrochloric acid. The precipitates were collected byfiltration, washed with water and dried over phosphorus pentoxide togive7-[2-(2-aminothiazol-4-yl)-2-allyloxyiminoacetamido]-3-[1-(2-aminoethyl)-1H-tetrazol-5-yl]thiomethyl-3-cephem-4-carboxylic acid (syn isomer, 0.3 g). The motherliquid and the aqueous washing solution were combined together andsubjected to column chromatography on macroporous nonionic adsorptionresin "Diaion HP-20" [trademark: Mitsubishi Chemical Industries Ltd.]and eluted with 10% aqueous isopropyl alcohol. The eluate wasconcentrated in vacuo and lyophilized to give the same object compound(0.8 g). Total yield 1.1 g.

I.R. ν_(max) ^(Nujol) : 3270, 3150, 1760, 1660, 1610^(cm-1)

N.M.R. δ(DMSO-d₆, ppm): 3.37 (2H, m), 3.60 (2H, m), 3.23 (2H, m),4.30-4.91 (4H, m), 4.91-5.50 (3H, m), 5.50-6.43 (2H, m), 6.69 (1H, s),7.17 (2H, broad s), 9.51 (1H, d, J=8.0 Hz)

EXAMPLE 16

(1) A mixture of 7-aminocephalosporanic acid (252.3 g.),1-(3-tert-butoxycarbonylaminopropyl)-1H-tetrazole-5-thiol (240 g.),sodium bicarbonate (171 g.), water (6 l.) and acetone (1.5 l.) wasstirred at 60° to 65° C. for 3 hours. The resultant mixture was cooledto 10° to 15° C. and adjusted to pH 4.0 with 10% hydrochloric acid. Theprecipitates were collected by filtration and washed with water andacetone in turn to give7-amino-3-[1-(3-tert-butoxycarbonylaminopropyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (189.4 g.).

I.R. ν_(max) ^(Nujol) : 3350, 3150, 1800, 1700, 1620, 1540 cm⁻¹

N.M.R. δ(Dcl+D₂ O, ppm): 1.25 (9H, s), 2.37 (2H, m), 3.23 (2H, m), 3.87(2H, s), 4.37 (2H, s), 4.67 (2H, t. J=7 Hz), 5.18 (1H, d, J=5 Hz), 5.37(1H, d, J=5 Hz)

(2) 2-(2-Formamidothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoaceticacid (syn isomer, 16.5 g.), N,N-dimethylformamide (4.4 g.), phosphorylchloride (9.2 g.),7-amino-3-[1-(3-tert-butoxycarbonylaminopropyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (23.6 g.), sodium carbonate (5.3 g.), tetrahydrofuran (100 ml.),acetone (150 ml.) and water (150 ml.) were treated in a similar mannerto that of Example 11-(1) to give7-[2-(2-formamidothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-[1-(3-tert-butoxycarbonylaminopropyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (syn isomer, 39.1 g.).

I.R. ν_(max) ^(Nujol) : 3250, 1780, 1680 (broad s), 1540 cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 1.40 (18H, s), 2.07 (2H, m), 2.97 (2H, m), 3.73(2H, m), 4.37 (4H, m), 4.67 (2H, s), 5.22 (1H, d, J=5 Hz), 5.88 (1H, dd,J=8 Hz, 5 Hz), 7.50 (1H, s), 8.57 (1H, s), 9.62 (1H, d, J=8 Hz)

(3) Conc. hydrochloric acid (20.8 g.) was added to a mixture of7-[2-(2-formamidothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-[1-(3-tert-butoxy-carbonylaminopropyl)-1H-tetrazol-5-ylthiomethyl)-3-cephem-4-carboxylicacid (syn isomer, 39.1 g.) in methanol (400 ml.), and stirred at roomtemperature for 4 hours. After removing the solvent in vacuo,tetrahydrofuran (40 ml.) and anisole (40 ml.) were added to the residueand stirred at room temperature for 4 hours. The reaction mixture wasadded to ethyl acetate at room temperature. The precipitates werecollected by filtration and washed with ethyl acetate to give7-[2-(2-aminothiazol-4-yl)-2-carboxymethoxyiminoacetamido]-3-[1-(3-aminopropyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (a mixture of syn and anti isomers). The product was subjected tocolumn chromatography on macroporous nonionic adsorption resin "HP-20"(trademark, manufactured by Mitsubishi Chemical Industries Co.) to givethe syn isomer of the objective compound.

I.R. _(max) ^(Nujol) : 3300, 1760, 1660, 1600, 1520 cm⁻¹

N.M.R. (DMSO-d₆, ppm): 2.34 (2H, m), 3.16 (2H, m), 3.64 (2H, q, J=17Hz), 4.24 (2H, q, J=13 Hz), 4.56-4.34 (4H, m), 5.20 (1H, d, J=5 Hz),5.74 (1H, d, J=5 Hz), 7.04 (1H, s)

EXAMPLE 17

Sodium bicarbonate (0.5 g.) was dissolved in pH 6.8 phosphate buffer (30ml.).7-[2-(2-Aminothiazol-4-yl)-2-propargyloxyiminoacetamido]-cephalosporanicacid (syn isomer, 1.5 g.) and1-(2-tert-butoxycarbonylaminoethyl)-1H-tetrazole-5-thiol (1.1 g.) wereadded to the solution at 60° to 65° C. and stirred at the sametemperature for 3 hours. Ethyl acetate and tetrahydrofuran (2:1) wereadded to the resultant mixture and adjusted to pH 3.0 with 10%hydrochloric acid. The organic layer was separated and the aqueous layerwas extracted with ethyl acetate. The ethyl acetate extract and theorganic layer were combined together, washed with a saturated sodiumchloride aqueous solution and dried over magnesium sulfate. Afterremoving the solvent in vacuo, the residue was dissolved in methanol (30ml.). After adding coc. hydrochloric acid (0.6 ml.) thereto, the mixturewas repeatedly evaporated by adding methanol. The residue was pulverizedwith diethyl ether and collected by filtration to give7-[2-(2-aminothiazol-4-yl)-2-propargyloxyiminoacetamido]-3-[1-(2-aminoethyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid dihydrochloride (syn isomer, 0.3 g.)

I.R. ν_(max) ^(Nujol) : 3300-3100, 1770, 1700 (sh), 1670, 1630, 1540cm⁻¹

N.M.R. δ(DMSO-d₆, ppm): 3.35 (2H, m), 3.55 (1H, m), 3.75 (2H, broad s),5.05-4.15 (4H, m), 4.78 (2H, m), 5.13 (1H, d, J=5 Hz), 5.73 (1H, dd, J=5Hz, 8 Hz), 6.93 (1H, s), 9.78 (1H, d, J=8 Hz)

EXAMPLE 18

4-Amino-5-methyl-3-mercapto-4H-1,2,4-triazole (1.5 g.) was added to astirred suspension of7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]cephalosporanic acid(syn isomer, 3.7 g.) in 0.2 M phosphate buffer (30.0 ml.) at 65° C., andadjusted to pH 6.5 to 6.8. The solution was stirred at 65° C. for 52/3hours. The resultant mixture was adjusted to pH 5.0 with 10%hydrochloric acid and subjected to column chromatography on macroporousnon-ionic adsorption resin "Diaion HP-20" (trademark: manufactured byMitsubishi Chemical Industries Co.) with 10-20% aqueous isopropylalcohol. The eluate was concentrated in vacuo and lyophilized to give7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-(4-amino-5-methyl-4H-1,2,4-triazol-3-ylthiomethyl)-3-cephem-4-4-carboxylicacid (syn isomer, 0.38 g.).

I.R. _(max) ^(Nujol) : 3350, 3250, 1770, 1670, 1600cm⁻¹

N.M.R. (DMSO-d₆, ppm): 2.25(3H, s), 3.45(2H, m), 3.79(3H, m), 4.15(2H,m), 4.91(1H, d, J=5 Hz), 5.53(1H, dd, J=5 Hz, 8 Hz), 5.83 (2H, broad s),6.69(1H, s), 7.16(2H, broad s), 9.72(1H, d, J=8 Hz).

EXAMPLE 19

(1)2-[2-(2,2,2-Trifluoroacetamido)thiazol-4-yl]-2-cyclopentyloxyiminoaceticacid (syn isomer, 1.5 g.), dry N, N-dimethylformamide (0.4 g.),phosphoryl chloride (0.8 g.),7-amino-3-[1-(3-tert-butoxycarbonylaminopropyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (2.0 g.), sodium bicarbonate (1.4 g.), water (12.0 ml.), acetone(12.0 g.), and dry ethyl acetate (0.4 g.) were treated in a similarmanner to that of Example 11-(1) to give7-[2-[2-(2,2,2-trifluoroacetamido)thiazol-4-yl]-2-cyclopentyloxyiminoacetamido]-3-[1-(3-tert-butoxycarbonylaminopropyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (syn isomer, 2.60 g.).

I.R. _(max) ^(Nujol) : 3180, 1780, 1680cm⁻¹

N.M.R. (DMSO-d₆, ppm): 1.25-2.26(10H, m), 1.41(9H, s), 2.97(2H, m), 3.74(2H, m), 4.06-4.57(4H, m), 4.78(1H, m), 5.21(1H, d, J=4.0 Hz), 5.87(1H,dd, J=4.0 Hz, 8.0 Hz), 7.53(1H, s), 9.70(1H, d, J=8.0 Hz).

(2)7-[2-[2-(2,2,2-Trifuoroacetamido)thiazol-4-yl]-2-cyclopentyloxyiminoacetamido]-3-[1-(3-tert-butoxycarbonylaminopropyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (syn isomer, 2.5 g.) was dissolved in a solution of conc.hydrochloric acid (0.65 g.) in methanol (20 ml.).

After evaporating methanol in vacuo, the residue was dissolved inmethanol (20 ml.) and evaporated in vacuo. Water (40 ml.) was added tothe residue and adjusted to pH 2.0 with sodium bicarbonate. To themixture were added sodium acetate tr hydrate (4.2 g.) andtetrahydrofuran (15 ml.), and stirred at room temperature overnight. Theresultant solution was concentrated in vacuo, and adjusted to pH 3.8with 1 N hydrochloric acid. The precipitates were collected byfiltration, washed with water and dried over magnesium sulfate to give7-[2-(2-aminothiazol-4-yl)-2-cyclopentyloxyiminoacetamido]-3-[1-(3-aminopropyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (syn isomer, 0.31 g.). The filtrate was subjected to columnchromatography on non-ionic macroporous adsorpotion resin "Diaion-HP-20"[trademark: manufactured by Mitsubishi Chemical Industries Co.] with15-20% aqueous isopropyl alcohol. The eluate was concentrated in vacuoand the residue was lyophilized to give the same object substance (0.62g.) total yield (0.93 g.).

I.R. _(max) ^(Nujol) : 3270, 3160, 1760, 1610cm⁻¹

N.M.R. (DMSO-d₆, ppm): 1.28-1.96(8H, m), 2.16(2H, m), 2.88(2H, m),3.54(2H, m), 4.12-4.81(5H, m), 5.02(1H, d, J=5.0 Hz), 5.67(1H, dd, J=5.0Hz, 8.0 Hz), 6.67 (1H, s), 9.40(1H, d, j=8.0 Hz)

What we claim is: 1.7-[2-(2-aminothiazol-4-yl)-2-allyloxyiminoacetamido]-3-[1-(2-aminoethyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (syn isomer). 2.7-[2-(2-aminothiazol-4-yl)-2-propargyloxyiminoacetamido]-3-[1-(2-aminoethyl)-1H-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylicacid (syn isomer), or its dihydrochloride.
 3. A pharmaceuticallyantibacterial composition comprising an effective amount of the compoundof claims 1 or 2 in association with a pharmaceutically acceptable,substantially non-toxic carrier or excipient.